Search published articles
Showing 48 results for Polymerase Chain Reaction
Mohammad Reza Arabestani , Mohammad Yousef Alikhani , Manoochehr Karami , Elham Salimi Ghale ,
Volume 73, Issue 12 (3-2016)
Volume 73, Issue 12 (3-2016)
Abstract
Background: Coagulase-negative staphylococci (CoNS) were considered as contaminats previously, but, during the past decade considered as one of the most common photogenic bacteria in hospital. Resistance to beta-lactams especially methicillin in staphylococcus species is being worrying in hospitals. Rapid identification of mechanisms of resistance and confirmation of their resistance to methicillin is a basic principle for antibiotic treatment. The aim of this study was to determine antibiotic resistance, frequency of mecA gene, and determination of SCCmec types in CoNS isolates from teaching hospitals in Iran.
Methods: The descriptive cross-sectional study was carried out one hundred clinical samples isolated from patients with an average age of 7-69 years at teaching hospitals in Hamadan City, Iran, from September 2014 to February 2015. After confirmation of isolates by microbiological standard biochemical tests, antimicrobial susceptibility testing was performed by disk agar diffusion (DAD) method. After extraction of isolated genomicm, mecA gene was detected. Then, the types of SCCmec were performed by PCR.
Results: In this study, 387 clinical samples were collected which among 100 CoNS isolated, Staphylococcus epidermidis was the most prevalent species with frequency 55 (55%), followed by S. haemolyticus 40(40%) and S. saprophyticus 5(5%). The highest antibiotic susceptibility was to rifampin 96(96%) and the lowest resistance was detected for trimethoprim/sulfamethoxazole (TMP/SMX) 47(47%). None of the strains were resistant to vancomycin. Resistance to methicillin was detected in 50% of CoNS isolates. Typing of SCCmec was performed by The polymerase chain reaction (PCR). Frequency types of SCCmec was type III with frequency 13(13%), type V 11(11%), type II 6(6%), type IV 4 (4%), type I 3(3%) respectively. Thirteen isolated was not typable in this study.
Conclusion: The result of this study showed that a large percentage of coagulase-negative staphylococci are resistance to methicillin, and the prevalence of SCCmec type was type III, which encodes the largest number of resistance genes. This information could be use in epidemiological study for preventing of infectious control in hospital and health centers.
Ehsan Sarraf Kazerooni , Ehteramolsadat Hosseini , Zohreh Sharifi , Azita Azarkeivan , Mehran Ghasemzadeh ,
Volume 74, Issue 5 (8-2016)
Volume 74, Issue 5 (8-2016)
Abstract
Background: Human leukocyte antigen E is a member of non-classical HLA class I. Interaction between HLA-E molecule on the target cells and inhibitory CD94/NKG2A receptor on the cell surface of natural killer (NK) cells has an important role in the regulation of immune system against pathogens; therefore different cell surface expression of HLA-E molecule plays an important role in host resistance against viral infections as well as host response to treatment. Considering this fact, we analyzed the frequency of different HLA-E genotypes (HLA-E*01010101, HLA-E*01030103, HLA-E*01010103) in major thalassemic patients who underwent frequent transfusion therapy and are thus more susceptible to infectious diseases.
Methods: This study was a cross-sectional study of 104 major thalassemic patients who referred to Tehran Thalassemia Clinic between the years 2015 to 2016. Blood DNA was extracted and proliferated by sequence-specific primer polymerase chain reaction (SSP PCR). The PCR product was subjected to electrophoresis on 1.5 percent agarose gel then DNA fragment bands on the gel were detected by exposing to UV light. Furthermore, PCR products were also subjected to sequencing analysis for further confirmation.
Results: From 104 patients in this study, 49 (47.1%) were man and 55 (52.9%) were women. These patients were in the age range of 16 to 43 years (mean+SD; 31.03±4.7 year). The frequency of HLA-E*01010103 genotype (64.4 percent) was significantly (P= 0.001) higher than the genotypes of HLA-E*01010101 (15.4%) and HLA-E*01030103 (20.2%) whereas there was no difference between the frequency of HLA-E*0103 allele (52.4%) and HLA-E*0101 (47.6%).
Conclusion: This is the first study that examined the HLA-E polymorphisms in Iranian thalassemic patients referred to Tehran Thalassemia Clinic. This study has shown that the frequency of HLA-E*01010103 genotype was significantly higher than other genotypes of HLA-E whereas there was no difference between the frequency of HLA-E*0103 allele and HLA-E*0101 allele. Whether different frequencies of HLA-E genotype may affect thalassemic patients’ susceptibility to blood-borne infections will be of interest for future studies.
Zahra Asghari Lalami, Ahmad Ebrahimi, Maryam-Sadat Daneshpour,
Volume 74, Issue 6 (9-2016)
Volume 74, Issue 6 (9-2016)
Abstract
Background: Obesity is one of the most important problems in developed countries and cause cardiovascular diseases, diabetes and hypertension. The complex phenotype influenced by both genetic and the environment factors. One of the most important genes which is effective in this phenotype is peroxisome proliferator-activated receptor gamma (PPAR-γ). This study was carried out of investigate the association of Pro12Ala (rs1801282) polymorphism in mentioned gene with obesity in Tehran Lipid and Glucose Study (TLGS).
Methods: The present study done in September 2014 in Cellular and Molecular Endocrine Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences. For the present case-control study 239 subjects with excess weight and body mass index more than 30 kg/m2 as a case and 240 subjects with normal weight and body mass index less than 25 kg/m2 as a control were selected. The rs1801282 was proliferated, detected and genotyped using tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method.
Results: The results indicated that there was significant association between the presence of risk allele G of rs1801282 and obesity disease in the TLGS population (P=0.000). Genotype and allelic frequencies of rs1801282 in patient and healthy group were: 55.2% and 23.8% for GG, 24.3% and 30.4% for GC, 20.5% and 45.8% for CC, 67% and 39% for G, 33% and 61% for C, respectively.
Conclusion: The results of study indicated that the presence of G allele could be increase 1.7 the risk of obesity. These differences in patient and healthy group lead us to select this marker as a genetic marker to predict the risk of obesity. There are statistical differences between the distribution of mentioned polymorphism in Tehranian population and other populations. However, replicating the study in a larger population of Tehranian people with more affected cases is suggested to generalize the results of this study.
Mohammad Khalili , Mohammad Reza Aflatoonian , Farzaneh Salari Aliabadi , Jalil Abshenas ,
Volume 74, Issue 7 (10-2016)
Volume 74, Issue 7 (10-2016)
Abstract
Background: Human brucellosis is a significant public health problem in many middle east countries including Iran. Brucella organisms, which are small aerobic, facultative intracellular coccobacilli, localize in the reproductive organs of host animals, causing abortions and sterility. They are shed in large numbers in the animal’s urine, milk, placental fluid, and other fluids. Dairy product from raw milk are a potential threat to public health in endemic developing countries. The gold standard for the diagnosis of brucellosis is isolation of Brucella species. However, isolation Brucella species is time consuming and needed to level 3 biocontainment facilities and highly skilled technical personnel to handle samples and live bacteria for eventual identification. Handling Brucella species increase risk of laboratory infection. Polymerase chain reaction (PCR) with high sensitivity and specifity overcomed to these disadvantages. The aim of this study was to detect Brucella species in milk from dairy cattle farms in Kerman province, Iran by PCR technique.
Methods: Forty and eight bulk tank milk (BTM) were collected from October 2015 to March 2016 from 48 dairy cattle farm including 4200 cows. DNA of milk samples extracted by lysis buffer and proteinase K method. All milk samples were examined by PCR to detect Brucella-specific DNA targeting IS 711. Positive samples must be showed 317 bp amplified, corresponding to the expected size of the IS 711 genome region in all Brucella species.
Results: Using IS711 primer were detected in 4 samples (8.3%) Brucella spp. from 48 BTM samples in this area.
Conclusion: The results indicate that brucellosis by Brucella species is endemic in the Kerman province dairy farms. Consumption of raw milk dairy products by individual farmers operating under poor hygienic conditions represents an high risk to public health. The need for implementing control measures and raising public awareness on zoonotic transmission of brucellosis are recommended. Vaccination of cattle is recommended for control of bovine brucellosis in enzootic areas with high prevalence rates.
Amir Mirzaie , Shoreh Zare Karizi ,
Volume 74, Issue 9 (12-2016)
Volume 74, Issue 9 (12-2016)
Abstract
Background: Centaurea cyanus is an endemic and well-known herbal medicine in Iran, is an annual flowering plant in the family of Asteraceae. The flowers are the part used in modern herbal medicine and are considered to have tonic, stimulant and emmenagogue properties, with action similar to that of blessed thistle. The aim this study was to investigate the phytochemical constituents of C. cyanus extract, its antioxidant, anti-tumor and anti-bacterial activities.
Methods: This experimental study was conducted from June to January of 2015 in Islamic Azad University of Varamin, Iran. At first, the phytochemical components of C. cyanus extract was analyzed using gas chromatography–mass spectrometry (GC-MS) method. Subsequently, the antibacterial potential of the extract was evaluated against 4 pathogenic bacteria including Staphylococcus aureus, Streptococcus pyogenes, Psedomonas aeroginosa and Klebsiella pnemoniae via minimum inhibitory concentration (MIC) mathod. Moreover, the anti-oxidant and anti-tumor activities of extract on colon cancer cell line (HT29) were investigate using DPPH and MTT colorimetric methods, respectively. Finally, the Bax and Bcl2 apoptosis gene expression level was analyzed by quantitative Real-time PCR technique.
Results: GC-MS analysis of C. cyanus extract was shown 19 major components and the most frequent component was belonged to n-Hexadecanoic acid (36.4%) and Linoleic acid (19.3%). The maximum antibacterial activity of extract was observed on S. aureus and P. aeroginosa isolates. The antioxidant activity of the extract was 0.109±0.07 mg/ml. Moreover, the MTT results show that extract had IC50= 26.04±0.45 on HT29 cell line. The Real-time PCR results showed the expression level of Bax and Bcl2 was significantly increased and decreased respectively in colon cancer cell line (2.63±0.54 (P< 0.05), 0.38±0.72 (P< 0.05)).
Conclusion: The results of this study show that the extract had significant anti-bacterial and anti-cancer effects and it appear that the extract has potential uses for pharmaceutical industries.
Association between 497A>G polymorphism in FCGR2A gene with recurrent miscarriage in infertile women
Zahra Mofidimanesh , Khadijeh Onsory , Anahita Mohseni Meybodi ,
Volume 74, Issue 11 (2-2017)
Volume 74, Issue 11 (2-2017)
Abstract
|
Background: The results indicated that the immunologic and genetic factors play a key role in the susceptibility to this syndrome compared to other risk factors. Immunoglobulin G, representing approximately 80% of Immunoglobulins in humans and the only way that IgG2 can be passed from mother to fetus blood circulation is binding to Fcgamma receptor (FcγR) classes which have been coded by Fcgamma receptor (FcγRIIA) gene. Any changes in the FcγRIIA gene structure such as mutations or polymorphisms can be considered as risk factors on the incidence of abortion through causing the inflammation or decreasing fetus safety. This receptor is the only which can have an interaction with IgG2 antibody and the Therefore, the current study was carried out to assess the association between R/H131 polymorphism in the FcγRIIA gene and susceptibility to recurrent abortions in Iranian women. Methods: For this reason, a case-control study was confirmed to compare the frequency of FCGR2A gene R/H131 polymorphism in 150 women with recurrent miscarriage history having normal karyotype and 150 healthy women with no abortion history as control which were collected in March 2014 up to September 2015, from Royan Institute for Reproductive, Tehran, Iran. The genomic DNA was extracted from peripheral blood leukocytes and genotyping was performed using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Results: The frequency of AA, AG, and GG genotypes in case and control groups were 31.3%, 54.7%, 14% and 27.3%, 49.2%, 23.5% respectively. According to the findings, the presence of the risk allele was not associated with increased risk of recurrent miscarriage compared with individuals lacking the risk allele and it statistically was significant (P= 0.11). No significant association was found between the age of participants and risk of abortion in Iranian studied population (P= 0.083). Conclusion: The results of present study do not support the previous findings of an association between R/H131 polymorphism in FCGR2A gene and recurrent miscarriage. |
Nazanin Talebabadi , Amirnader Emami-Razavi, Raheleh Safaei-Javan, Hadis Mohammadpour , Alireza Abdollahi ,
Volume 75, Issue 1 (4-2017)
Volume 75, Issue 1 (4-2017)
Abstract
Background: As far as the role and amount of Transferrin receptor 2 (TFR2), which is the transferrin receptor gene, studies have been conducted, some of which confirming its relationship with gastric adenocarcinoma. The idea behind this study was to examine changes in the TFR2 gene expression in the tumor cells of gastric adenocarcinoma and comparing with gene expression in the normal tissue adjoining the tumor.
Methods: This case-control study was conducted at the Pathology Section of Cancer Institute of Imam Khomeini University Hospital in Tehran from September 2015 to September 2016. In this study, 30 fresh samples from tumor tissues of patients diagnosed with gastric adenocarcinoma, 30 fresh samples of normal tissue adjoining the tumor and 30 samples of frozen plasma from the same patients were taken. The patients' plasma was examined in terms of existence of helicobacter pylori antibody by enzyme linked immunosorbent assays (ELISA) method and TFR2 gene expression in the tumor tissue and the adjoining normal tissue by applying real-time polymerase chain reaction (Real-Time PCR).
Results: Gene expression (by applying real time polymerase chain reaction) in the tumor tissue was meaningfully higher than in the normal tissue (P= 0.125). The TFR2 expression in patients with stomach cancer, who were at the same time infected with helicobacter pylori, indicated that the gene expression had increased in those with this contamination (P= 0.077). Examining the relationship between this gene expression and the stage of disease showed that the TFR2 gene expression increased significantly in the more advanced stages of the disease (P= 0.396).
|
Conclusion: The TFR2 gene expression increases in the stomach's tumor tissue. This gene expression is higher in people infected with helicobacter pylori or in those at an advanced stage of the disease. These findings may confirm the direct relationship between gene expression and the occurrence or metastasis of gastric adenocarcinoma. |
Sadegh Baniaghil, Gholamreza Nikbakht Borujeni , Hassan Tajbakhsh, Atefeh Esmailnejad, Ali Akbar Amirzargar ,
Volume 75, Issue 3 (6-2017)
Volume 75, Issue 3 (6-2017)
Abstract
Background: HLA disease association was investigated in several autoimmune, cancer and infectious diseases. The outcome of tuberculosis (TB) infection may be influenced by host genetic factors like MMP-1, MCP-1, IL-10, IL-12, TNF-α, IFN-γ and human leukocyte antigen (HLA). Given the paucity of information with regard to the association between the human leukocyte antigens (HLA) and TB infection among Iranians, we aimed to identify HLA polymorphisms that might confer susceptibility or protect against TB.
|
Methods: In this case-control study, to investigate the association between the HLA-DRB1 and DQB1 alleles and TB, 50 patients with tuberculosis were selected from Sistani population in Golstan University of Medical Sciences, Golestan Province, North East of Iran, from September 2015 to February 2016. Allele frequencies in patients were compared with a 100 aged and sex match control group from healthy blood donor of that ethnic population. HLA-DRB1 and -DQB1 alleles were determined using polymerase chain reaction based on sequence specific primer (PCR-SSP) method by low to intermediate resolution kits supplied by CTS (Collaborative Transplant Study, Heidelber University, Germany). Using EPI-info statistical software Chi-square test and fisher exact test, 95% confidence interval and odd ratio were calculated and allele frequencies in patients and control subjects were compared. P-value less than 0.05 were considering statistically significant. |
Results: The results of this study showed a significant increase and positive association with -DRB1*04:03 (OR=3.13, CI 95% (2.47-3.96), -DRB1*14:04 (OR=3.13, CI 95% (2.47-3.96), -DQB1*0201 (OR=2.67, CI 95% (1.18-6.04), -DQB1*0601 (OR=3.16, CI 95% (1.36-7.73) ,while the frequency of -DRB1*07 (OR=0.16, CI 95% (0.05-0.52) were lower in patients than control group and shows negative association.
|
Conclusion: The results of this study confirmed some of the previous positive and/or negative association, however it is suggested that HLA-DRB1*04:03, -DRB1*14:04, -DQB1*0201, -DQB1*0601- have an important role in susceptibility to tuberculosis infection and -DRB1*07 was associated with protection in Iranian Sistani population. Larger case-control sample size studies may be helpful to confirm our investigation. In addition population-specific studies is needed for evaluation of the role of HLA polymorphisms in tuberculosis in different ethnic groups. |
Mohammad Mehdi Soltan Dallal, Shirin Nezamabadi, Jalal Mardaneh, Zahra Rajabi, Abolfazl Sirdani,
Volume 75, Issue 3 (6-2017)
Volume 75, Issue 3 (6-2017)
Abstract
Background: In recent years, use of powdered infant formula (PIF) milk for neonates feed is increasing; therefore, the quality control (QC) of PIF products is very important. The aim of present study was detection of toxigenic Bacillus cereus species in PIF milk using PCR assay.
Methods: The cross-sectional study was carried out on 125 samples of powdered infant formula milk (PIF) purchased between March 2015 and April 2016 in Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Briefly, 0.1 dilutions were prepared and inoculated on Bacillus cereus selective media (MYP) and incubated at 30 °C for 24 hours. The suspicious colonies were verified using biochemical tests based on standard methods. Final confirmation of studied isolates was carried out by ITS gene detection using polymerase chain reaction (PCR) assay. Presence of nonhemolytic enterotoxin (NHE) (linked to diarrhoea syndrome) and emetic toxin (EM) (linked to emetic syndrome) virulence genes were investigated using polymerase chain reaction assay.
|
Results: In this study, of 125 PIF samples, 84 (67.2%) were contaminated. Of various recovered bacteria from these samples, 110 bacterial isolates were suspected to be Bacillus spp. using phenotypic methods. The ITS PCR results showed that 91.8% of the isolates were B. cereus. Respectively, 53.63 and 79% of B. cereus isolates possessed NHE and EM virulence genes. |
Conclusion: Our data revealed that near 80% of Bacillus cereus isolates have emetic toxin (EM) gene, as result virulence potency of this isolates is very high. However, the low number of this organisms in foods is very important and food safety protocols for these opportunistic toxigenic bacteria should be revised. Since the pasteurization process is ineffective on B. cereus spores; therefore, spores can remain in PIF milk and the vegetative bacterial cells can cause food poisoning in neonates. Therefore, modification of foods quality control protocols is essential in order to identify virulence genes in this bacterium.
Amir Mirzaie , Aliasghar Bagheri Kashtali , Hassan Sahebjamee , Hassan Noorbazargan , Hassan Rahmati , Seyed Ataollah Sadat Shandiz,
Volume 75, Issue 5 (8-2017)
Volume 75, Issue 5 (8-2017)
Abstract
Background: Medicinal plants have been identified and used from prehistoric times and these plants make many chemical compounds for biological functions. Trifolium cherleri is an herbaceous species belonging to the family of the Fabaceae to Africa, Eurasia and Australia. T. cherleri is an important member of the Fabaceae family that is well-known herbal medicine in Iran. The aim of this study was to investigate the phytochemical composition, antibacterial and anti-cancer activities of T. cherleri extract.
Methods: This experimental study was performed in Islamic Azad University, from December 2016 to February 2017. At first, the phytochemical constituents of T. cherleri extract were determined using gas chromatography-mass spectrometry (GC-MS) method. Subsequently, the antibacterial activity of the extract was evaluated against some gram positive and negative pathogenic bacteria included Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC 19615, Salmonella enteritidis ATCC 13076 and Listeria monocytogenes ATCC 35152 via minimum inhibitory concentration (MIC) method. Moreover, anticancer potential of extract was examined by colorimetric MTT assay toward lung cancer (A549) cell line. Then, the evaluation of caspase 3 and 9 apoptosis gene expression was determined using Real-Time Polymerase Chain Reaction (Real-Time PCR) technique. Moreover, the Real-Time PCR was performed using relative quantitative method.
Methods: This experimental study was performed in Islamic Azad University, from December 2016 to February 2017. At first, the phytochemical constituents of T. cherleri extract were determined using gas chromatography-mass spectrometry (GC-MS) method. Subsequently, the antibacterial activity of the extract was evaluated against some gram positive and negative pathogenic bacteria included Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC 19615, Salmonella enteritidis ATCC 13076 and Listeria monocytogenes ATCC 35152 via minimum inhibitory concentration (MIC) method. Moreover, anticancer potential of extract was examined by colorimetric MTT assay toward lung cancer (A549) cell line. Then, the evaluation of caspase 3 and 9 apoptosis gene expression was determined using Real-Time Polymerase Chain Reaction (Real-Time PCR) technique. Moreover, the Real-Time PCR was performed using relative quantitative method.
| Results: The phytochemical analyses of T. cherleri extract showed the 20 major components and the most frequent component was belonged to hexadecanoic acid, ethyl ester (20.7%) and 2-Pentadecanone, 6,10,14-trimethyl (19.9%). The extract had maximum antibacterial effects against Staphylococcus aureus and Streptococcus pyogenes. There was a dose dependent increase in the cytotoxicity effect of extract against A549 cancer cell. Moreover, the Real-Time PCR results indicated that the caspase 3 and caspase 9 gene expression was significantly up-regulated 2.57±0.27 (P<0.05), and 3.3±0.46 (P<0.05), respectively. |
Conclusion: The results of this study showed that the T. cherleri extract had significant anti-bacterial and anti-cancer effects and it appear that the extract has potential uses for pharmaceutical industries. Moreover, it could be considered as a promising source for novel drug compounds, but more studies are needed.
Neda Norouzi , Mortaza Bonyadi , Esmaeil Babaei , Mohammad Hossein Jabbarpour Bonyadi ,
Volume 75, Issue 5 (8-2017)
Volume 75, Issue 5 (8-2017)
Abstract
Background: Age-related macular degeneration (AMD) is the leading cause of blindness in the developed world and is characterized by progressive degeneration of the retinal pigment epithelium and secondary photoreceptor loss, resulting in visual loss. Etiological research suggests that age related macular degeneration is a complex disease, caused by the interactions of several genetic and environmental factors. Polymorphisms in genes encoding the alternative complement pathway, complement factor I (CFI), are associated with the risk for age related macular degeneration. The purpose of this investigation was studying of complement factor I p.Gly119Arg (C.355G>A) polymorphism with age related macular degeneration in the population living in Tehran, Iran.
Methods: This case-control study was conducted at Tabriz University from June 2015 to June 2016. In this study the association of p.Gly119Arg polymorphism in complement factor I gene was investigated in 150 patients suffering from age-related macular degeneration and 150 healthy age, sex and ethnicity matched unrelated people as control group. Both of the case and control groups were originated from the population living in Tehran. Genotypes of both groups were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and data was analyzed the Chi-square test in 2x2.Contingency table.
Methods: This case-control study was conducted at Tabriz University from June 2015 to June 2016. In this study the association of p.Gly119Arg polymorphism in complement factor I gene was investigated in 150 patients suffering from age-related macular degeneration and 150 healthy age, sex and ethnicity matched unrelated people as control group. Both of the case and control groups were originated from the population living in Tehran. Genotypes of both groups were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and data was analyzed the Chi-square test in 2x2.Contingency table.
| Results: Investigation of the association of p.Gly119Arg polymorphism in complement factor I gene with age related macular degeneration showed that there are statistically significant differences between patients and controls in genotype and allele frequencies of this polymorphism (P=0.005 and OR=6.68 in TT, P=0.04 and OR=0.61 in CC, P=0.03 and OR=1.76 in T, P=0.04 and OR=0.56 in C). Therefore CC, TT genotypes and C, T alleles were significantly associated with age related macular degeneration. |
Conclusion: This study showed a significant association between this polymorphism p.Gly119Arg (C.355G>A) complement factor I gene and age related macular degeneration disease in the population living in Tehran (P<0.05). Our data suggests that this locus polymorphism is not as rare in our studied population as previously reported from different population.
Maryam Khanmohamadi , Amir Seyed Ali Mehbod , Mojtaba Noraeepour , Mojtaba Didehdar ,
Volume 75, Issue 7 (10-2017)
Volume 75, Issue 7 (10-2017)
Abstract
Background: Vulvovaginal candidiasis (VVC) is a common infection, affecting up to 75% of women during their lifetimes. Approximately 5% of patients may experience recurrent VVC. Candida albicans is the most common causative agent of VVC. The objectives of this study were identification of candida species isolated of women with vulvovaginal candidiasis by molecular method in Arak city.
Methods: In this descriptive cross-sectional study, between Jun 2015 to March 2016 from 210 patients with vulvovaginal candidiasis referred to gynecology and obstetrics clinics in Arak city, Iran. Vaginal sampling was performed by wet sterile swabs. Samples were collected from vaginal discharge, vaginal posterior fornix, and sides of the vaginal wall. The swabs were investigated for direct exam and cultured on Sabouraud’s dextrose agar medium contain chloramphenicol. Yeast isolates DNA were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Fungal genomic DNA was extracted from each isolate colony, glass bead method and after amplification of ITS1-ITS4 region with PCR assay, digested by MSP I restriction enzyme.
Results: From 210 patients with vulvovaginitis, 95 (45.2%) patients showed VVC. These patients were positive for Candida growth in culture and were infected with one Candida species. The age range of women with vulvovaginitis was between 14-60 years and the most VVC cases were in age group of 21-30 years. The most common Candida species isolated were Candida. albicans (70.5%), C. glabrata (20%), C. tropicalis (7.4%) and C. parapsilosis (2.1%).
Conclusion: Regarding to the results of this study, C. albicans was the most common Candida species, isolated from patients with vulvovaginal candidiasis and approximately 30% of this infection causing by non-albicans species of Candida.
Methods: In this descriptive cross-sectional study, between Jun 2015 to March 2016 from 210 patients with vulvovaginal candidiasis referred to gynecology and obstetrics clinics in Arak city, Iran. Vaginal sampling was performed by wet sterile swabs. Samples were collected from vaginal discharge, vaginal posterior fornix, and sides of the vaginal wall. The swabs were investigated for direct exam and cultured on Sabouraud’s dextrose agar medium contain chloramphenicol. Yeast isolates DNA were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Fungal genomic DNA was extracted from each isolate colony, glass bead method and after amplification of ITS1-ITS4 region with PCR assay, digested by MSP I restriction enzyme.
Results: From 210 patients with vulvovaginitis, 95 (45.2%) patients showed VVC. These patients were positive for Candida growth in culture and were infected with one Candida species. The age range of women with vulvovaginitis was between 14-60 years and the most VVC cases were in age group of 21-30 years. The most common Candida species isolated were Candida. albicans (70.5%), C. glabrata (20%), C. tropicalis (7.4%) and C. parapsilosis (2.1%).
Conclusion: Regarding to the results of this study, C. albicans was the most common Candida species, isolated from patients with vulvovaginal candidiasis and approximately 30% of this infection causing by non-albicans species of Candida.
Hoda Ahmadi , Reza Mirfakhraie , Shiva Irani ,
Volume 75, Issue 10 (1-2018)
Volume 75, Issue 10 (1-2018)
Abstract
Background: Recurrent pregnancy loss is a form of infertility with at least three consecutive pregnancy losses or more. Y chromosome microdeletions are a class of most likely genetic factors that occur in a special zone of Y chromosome which is named azoospermia factor region. The purpose of this study was to analyze the presence of Y chromosome complete microdeletions in male partner of couples suffering from idiopathic recurrent pregnancy loss among Iranian population.
Methods: In the present study, Y chromosome microdeletions were evaluated in ninety-two male partners of couples with the experience of recurrent pregnancy loss as the patient group and also a group containing fifty fertile males as the control group. The research has done in Medical Genetic laboratory of Tehran and Islamic Azad University Science and Research Branch, Tehran, Iran within June 2013 to September 2014. The selected sequence tagged site markers (primers) including sY84, sY86, for azoospermia factor a; sY127, sY134, sY129, for azoospermia factor b and sY254, sY255, for azoospermia factor c were used to screen complete microdeletions in Y chromosome. At the first step DNA samples were extracted from all men’s peripheral blood in both patient and control groups and then multiplex polymerase chain reaction and also agarose gel electrophoresis were performed on this DNA samples so as to detect deletions.
Results: With due attention to the data resulted from multiplex polymerase chain reaction and agarose gel electrophoresis in order to recognize Y chromosome micro deletions in azoospermia factor region, in this work, all the bands related to the mentioned primers which were formed during the polymerase chain reaction, were detected on the gel obviously. It means that none of the samples neither the fifty fertile men nor the ninety-two patient men had complete micro deletions in their Y chromosome.
Conclusion: This study suggests that there is no correlation between Y chromosome micro deletions and occurrence of recurrent pregnancy loss in Iranian population.
Methods: In the present study, Y chromosome microdeletions were evaluated in ninety-two male partners of couples with the experience of recurrent pregnancy loss as the patient group and also a group containing fifty fertile males as the control group. The research has done in Medical Genetic laboratory of Tehran and Islamic Azad University Science and Research Branch, Tehran, Iran within June 2013 to September 2014. The selected sequence tagged site markers (primers) including sY84, sY86, for azoospermia factor a; sY127, sY134, sY129, for azoospermia factor b and sY254, sY255, for azoospermia factor c were used to screen complete microdeletions in Y chromosome. At the first step DNA samples were extracted from all men’s peripheral blood in both patient and control groups and then multiplex polymerase chain reaction and also agarose gel electrophoresis were performed on this DNA samples so as to detect deletions.
Results: With due attention to the data resulted from multiplex polymerase chain reaction and agarose gel electrophoresis in order to recognize Y chromosome micro deletions in azoospermia factor region, in this work, all the bands related to the mentioned primers which were formed during the polymerase chain reaction, were detected on the gel obviously. It means that none of the samples neither the fifty fertile men nor the ninety-two patient men had complete micro deletions in their Y chromosome.
Conclusion: This study suggests that there is no correlation between Y chromosome micro deletions and occurrence of recurrent pregnancy loss in Iranian population.
Sajad Shafai , Elham Moslemi , Mehdi Mohammadi , Kasra Esfahani , Amir Izadi ,
Volume 75, Issue 10 (1-2018)
Volume 75, Issue 10 (1-2018)
Abstract
Background: Prostate cancer is one of the most common diseases that affect men. Although prostate cancer is not the fatal flaw in most cases, detection of effective factors for early diagnosis and treatment is essential. Research results have shown that the use of KLK2 plus PSA can be a good biomarker for diagnosing prostate cancer. During prostate cancer, expression of KLK2 gene increases which can be used as a prostate cancer biomarker. The aim of this study is an assessment of KLK2 gene expression as a potential factor in the prostate cancer diagnosis.
Methods: In this case study, 50 prostate cancer urine samples from patients and 50 urine samples from normal individuals who were referred to Mehr Hospital of Tehran (from December 2014 to February 2016) were obtained and stored in the central research laboratory of Shahid Beheshti University of Medical Sciences, Tehran, till tests were being done. The age of collected samples between the 46 up to 71 years. RNA of samples were extracted, and then cDNA was synthesized by using M-MuLV enzyme, Oligo dt, and Random hexamer primers. KLK2 specific primers designed by Primer Express software, version 3.0 (Applied Biosystems, Foster City, CA, USA), and KLK2 gene expression evaluated by using ∆∆ct methods.
Results: In comparison with patients and normal sample`s gene expression, the mean increase expression of KLK2 gene in patients less than 50 years was 2.32 and in patients more than 50 years, it was 5.79, P<0.0001. In addition, gene expression results with respect to GS (Gleason grading system) classification shown that patients with GS6 had the lowest gene expression (3.40) and in the patients with GS8, had the highest gene expression (10.74) in comparison with normal group (P<0.0001).
Conclusion: The expression of KLK2 gene in people with prostate cancer is the higher than the healthy person; finally, according to the results, it could be mentioned that the KLK2 gene considered as a useful factor in prostate cancer, whose expression is associated with progression and development of the prostate cancer.
Methods: In this case study, 50 prostate cancer urine samples from patients and 50 urine samples from normal individuals who were referred to Mehr Hospital of Tehran (from December 2014 to February 2016) were obtained and stored in the central research laboratory of Shahid Beheshti University of Medical Sciences, Tehran, till tests were being done. The age of collected samples between the 46 up to 71 years. RNA of samples were extracted, and then cDNA was synthesized by using M-MuLV enzyme, Oligo dt, and Random hexamer primers. KLK2 specific primers designed by Primer Express software, version 3.0 (Applied Biosystems, Foster City, CA, USA), and KLK2 gene expression evaluated by using ∆∆ct methods.
Results: In comparison with patients and normal sample`s gene expression, the mean increase expression of KLK2 gene in patients less than 50 years was 2.32 and in patients more than 50 years, it was 5.79, P<0.0001. In addition, gene expression results with respect to GS (Gleason grading system) classification shown that patients with GS6 had the lowest gene expression (3.40) and in the patients with GS8, had the highest gene expression (10.74) in comparison with normal group (P<0.0001).
Conclusion: The expression of KLK2 gene in people with prostate cancer is the higher than the healthy person; finally, according to the results, it could be mentioned that the KLK2 gene considered as a useful factor in prostate cancer, whose expression is associated with progression and development of the prostate cancer.
Nahid Arefi Lisar , Parivash Kordbacheh , Sasan Rezaie , Mahin Safara , Roshanak Daie Ghazvini , Heidar Bakhshi , Zahra Omidvar Jalali ,
Volume 75, Issue 12 (3-2018)
Volume 75, Issue 12 (3-2018)
Abstract
Background: Vaginal candidiasis is common in during pregnancy. It may lead to complications like abortions, premature birth, low birth weight, chorioamnionitis and fungal systemic neonatal infection. The aim of present study was identification of Candida species by mycological and molecular methods in pregnant women with vaginal candidiasis.
Methods: This cross-sectional study was performed on 80 pregnant women with or without clinical symptoms of vulvovaginal candidiasis referred to Shahid Noorani Talesh Hospital, Gilan University of Medical Sciences, Iran, from April to December 2015 (8 months). All specimens were examined by direct microscopy and culture on CHROMagar Candida medium for isolation and differentiation of major clinical-significant Candida species (spp.). Cultured media were incubated at 35 °C for 48 hours and evaluated based on color and number of grown colonies. If no growth was observed, the media were incubated for several additional days. Subcultures were done on Sabouraud dextrose agar (Merck, Germany) and Corn meal agar with Tween 80 media (Micromedia, Hungary) for further study. Identification of Candida spp. carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.
Results: In this study, vulvovaginal candidiasis was observed in 20 (25%) patients. Twenty-two isolates were obtained from culture of specimens on CHROMagar Candida medium (Paris, France). The most common isolated species was Candida albicans 16 (72.8%) and followed by Candida glabrata 5 (22.7%), Candida tropicalis 3 (13.6%) and Candida krusei 1 (4.5%) cases. Two patients had mixed infection with 2 different Candida species (C. albicans and C. glabrata) While using PCR-RFLP method, the Candida species were identified as 13 (59.1%) Candida albicans, 5 (22.7%) Candida glabra, 3 (13.6%) Candida tropicalis and 1 (4.5%) Candida krusei cases, respectively. In direct examination were seen yeast budding cells and pseudohyphae in 8 culture positive specimens. In the present study, results of conventional mycological method in differentiation of Candida spp. were consistent with molecular results in 80% of cases. There was also significant correlation between vulvovaginal candidiasis with clinical symptoms (P<0.0001), including diabetes mellitus (P<0.014), and taking antibacterial drugs (P<0.003) in pregnant women.
Conclusion: PCR-RFLP was able to identify correctly the Candida spp. as a complementary method.
Methods: This cross-sectional study was performed on 80 pregnant women with or without clinical symptoms of vulvovaginal candidiasis referred to Shahid Noorani Talesh Hospital, Gilan University of Medical Sciences, Iran, from April to December 2015 (8 months). All specimens were examined by direct microscopy and culture on CHROMagar Candida medium for isolation and differentiation of major clinical-significant Candida species (spp.). Cultured media were incubated at 35 °C for 48 hours and evaluated based on color and number of grown colonies. If no growth was observed, the media were incubated for several additional days. Subcultures were done on Sabouraud dextrose agar (Merck, Germany) and Corn meal agar with Tween 80 media (Micromedia, Hungary) for further study. Identification of Candida spp. carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.
Results: In this study, vulvovaginal candidiasis was observed in 20 (25%) patients. Twenty-two isolates were obtained from culture of specimens on CHROMagar Candida medium (Paris, France). The most common isolated species was Candida albicans 16 (72.8%) and followed by Candida glabrata 5 (22.7%), Candida tropicalis 3 (13.6%) and Candida krusei 1 (4.5%) cases. Two patients had mixed infection with 2 different Candida species (C. albicans and C. glabrata) While using PCR-RFLP method, the Candida species were identified as 13 (59.1%) Candida albicans, 5 (22.7%) Candida glabra, 3 (13.6%) Candida tropicalis and 1 (4.5%) Candida krusei cases, respectively. In direct examination were seen yeast budding cells and pseudohyphae in 8 culture positive specimens. In the present study, results of conventional mycological method in differentiation of Candida spp. were consistent with molecular results in 80% of cases. There was also significant correlation between vulvovaginal candidiasis with clinical symptoms (P<0.0001), including diabetes mellitus (P<0.014), and taking antibacterial drugs (P<0.003) in pregnant women.
Conclusion: PCR-RFLP was able to identify correctly the Candida spp. as a complementary method.
Seyedeh Hakimeh Rezazadeh, Reza Shirkoohi, Abdolhamid Angaji, Seyed Yusef Seyedena, Amir Nader Emami Razavi,
Volume 76, Issue 2 (5-2018)
Volume 76, Issue 2 (5-2018)
Abstract
Background: Ovarian cancer is a leading metastatic disease. The epithelial ovarian cancer is one of the most common malignant cancers that usually remains asymptomatic up to metastasis stages, and most patient when diagnosed are in the advanced stage of the disease. Studies have shown that in the majority of epithelial cancers mesenchymal factor expression such as Vimentin increases, and the epithelial factor expression such as E-cadherin decreases, as a result, it causes an epithelial-mesenchymal transition (EMT). The aim of this study was to determine the expression level of these genes and association between EMT phenomenon and development of ovarian cancer based on clinical and morphological findings.
Methods: In the present case series study, 70 samples were chosen from the tumor Bank of Cancer Institute taken from patients at Imam Khomeini Hospital, Tehran, Iran. The amount of expression of two genes, E-cadherin and vimentin, was investigated by real-time PCR method from February 2016 to September 2017. The RNA extraction was done manually, and then cDNA synthesis was performed; In each sample the expression level of vimentin and E-cadherin was measured with real-time PCR method. The patient’s clinical information with other data were analyzed with nonparametric statistical methods in SPSS software, version 19 (SPSS Inc., Chicago, IL, USA).
Results: There was a significant relationship between expression of vimentin gene and the stage (P=0.026) of the disease and metastasis (P=0.009), There was no significant relationship between vimentin gene expression and tumor grade (P=0.207), age (P=0.11), tumor size (P=0.71) and family history (P=0.6). There was a significant correlation between E-cadherin gene expression and metastasis (P=0.027), no significant correlation was found between E-cadherin gene expression with tumor grade (P=0.690), stage (P=0.753), age (P=0.09), tumor size (P=0.537) and family history (P=0.56).
Conclusion: According to the changes in expression of vimentin and E-cadherin genes in ovarian tumor cells, and association between these two genes with clinical and morphological findings and the role of these genes in the migration and invasion, we can use the both genes, vimentin and E-cadherin, as genes involved in the EMT process to assess disease progression and incidence of cell invasion in ovarian cancer.
Methods: In the present case series study, 70 samples were chosen from the tumor Bank of Cancer Institute taken from patients at Imam Khomeini Hospital, Tehran, Iran. The amount of expression of two genes, E-cadherin and vimentin, was investigated by real-time PCR method from February 2016 to September 2017. The RNA extraction was done manually, and then cDNA synthesis was performed; In each sample the expression level of vimentin and E-cadherin was measured with real-time PCR method. The patient’s clinical information with other data were analyzed with nonparametric statistical methods in SPSS software, version 19 (SPSS Inc., Chicago, IL, USA).
Results: There was a significant relationship between expression of vimentin gene and the stage (P=0.026) of the disease and metastasis (P=0.009), There was no significant relationship between vimentin gene expression and tumor grade (P=0.207), age (P=0.11), tumor size (P=0.71) and family history (P=0.6). There was a significant correlation between E-cadherin gene expression and metastasis (P=0.027), no significant correlation was found between E-cadherin gene expression with tumor grade (P=0.690), stage (P=0.753), age (P=0.09), tumor size (P=0.537) and family history (P=0.56).
Conclusion: According to the changes in expression of vimentin and E-cadherin genes in ovarian tumor cells, and association between these two genes with clinical and morphological findings and the role of these genes in the migration and invasion, we can use the both genes, vimentin and E-cadherin, as genes involved in the EMT process to assess disease progression and incidence of cell invasion in ovarian cancer.
Leila Hassani , Golnaz Asaadi Tehrani , Sina Mirza Ahmadi ,
Volume 76, Issue 7 (10-2018)
Volume 76, Issue 7 (10-2018)
Abstract
Background: Glioma is one of the most common and deadliest primary malignant tumors in the brain. A large part of the gene expression products are non-coding protein RNA. LncRNA THRIL gene is an antisense LncRNA and one of the most important mediators of the NF-KB signaling pathway, that express in many tissues of the body, including the central nerve system (CNS). The aim of the present study was to investigate the alternation in the expression of LncRNA THRIL gene in cells of the adenocarcinoma glioblastoma T98G cell line, under treatment with temozolomide chemotherapy drug.
Methods: This case-control study was conducted in Research Center of Islamic Azad University of Zanjan, Iran, from April to September 2017. Cells of T98G cell line was treated with various doses of temozolomide chemotherapy drug (25-50-100 μM) and at different times (72-48-48 hours), Respectively RNA extraction and cDNA synthesis were performed. Then LncRNA THRIL gene expression was studied by real-time PCR method and the results were analyzed by relative quantitative and livak methods.
Results: The THRIL gene expression in 24 hours’ time with 25 and 100 μM doses (P< 0.001) had significant decreased expression and 50 μM dose had non-significant increase. It had significant increase in 48 hours time with 50 μM dose (P< 0.001), except 25 μM (P< 0.001) and 100 μM (P< 0.001) doses had significant decreased expression. It had significant increase during 72 hours time with 50 μM dose (P< 0.001), in contrast 25 μM (P> 0.001) and 100 μM (P< 0.001) concentrations of temozolomide chemotherapy drug demonstrated decreased expression of the LncRNA THRIL gene (P< 0.001).
Conclusion: As a result, the THRIL gene expression alterations after cancer cells treatment by temozolomide chemotherapy drug depends on the time and dose of the drug and the 48 hours treatment time with 50 μM dose (P< 0.001) had the highest effect on cancer cells of the T98G cell line, due to the expression of the THRIL gene.
Methods: This case-control study was conducted in Research Center of Islamic Azad University of Zanjan, Iran, from April to September 2017. Cells of T98G cell line was treated with various doses of temozolomide chemotherapy drug (25-50-100 μM) and at different times (72-48-48 hours), Respectively RNA extraction and cDNA synthesis were performed. Then LncRNA THRIL gene expression was studied by real-time PCR method and the results were analyzed by relative quantitative and livak methods.
Results: The THRIL gene expression in 24 hours’ time with 25 and 100 μM doses (P< 0.001) had significant decreased expression and 50 μM dose had non-significant increase. It had significant increase in 48 hours time with 50 μM dose (P< 0.001), except 25 μM (P< 0.001) and 100 μM (P< 0.001) doses had significant decreased expression. It had significant increase during 72 hours time with 50 μM dose (P< 0.001), in contrast 25 μM (P> 0.001) and 100 μM (P< 0.001) concentrations of temozolomide chemotherapy drug demonstrated decreased expression of the LncRNA THRIL gene (P< 0.001).
Conclusion: As a result, the THRIL gene expression alterations after cancer cells treatment by temozolomide chemotherapy drug depends on the time and dose of the drug and the 48 hours treatment time with 50 μM dose (P< 0.001) had the highest effect on cancer cells of the T98G cell line, due to the expression of the THRIL gene.
Milad Pezeshki , Jamshid Ansari ,
Volume 76, Issue 10 (1-2019)
Volume 76, Issue 10 (1-2019)
Abstract
Background: Breast cancer is one of the most common worldwide malignancies among women. Biological data suggest that damage induced by endogenous and exogenous factors affects the integrity of DNA and associated with susceptibility to breast cancer. Single nucleotide polymorphisms (SNPs) in DNA repair genes can associated with differences in the repair efficiency of DNA damage and may affect breast cancer. The XRCC3 protein participates in DNA double-strand breaks and recombinational repair, in other words the product of XRCC3 gene, plays a key role in homologous recombination repair of DNA double-strand breaks. The polymorphism of FokI plays critical roles in breast cancer development. The aim of the present study was to evaluate associations between the risk of breast cancer and FokI polymorphism in the XRCC3 gene.
Methods: This case-control study was carried out on the women with breast cancer and healthy women located in Markazi province at Arak University Research, Iran, from October 2016 to March 2017. In the present study, the association of FokI polymorphisms and the risk of breast cancer was assessed by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques. In this method, genomic DNA was extracted from blood samples using the kit procedure. Then, PCR was performed and the SNP-containing DNA amplicons were subjected to digestion of enzymes. Following digestion, each sample was immediately analyzed by 3% agarose gel electrophoresis. Statistical analysis was done using SPSS and SNP Analyzer softwares and the final results were determined.
Results: No statistically significant difference was observed between the two groups of patients and controls for three genotypes the site rs1799794 (P=0.435). Genotype AG (P=0.384, OR=0.614, CI=95%, 0.205-1.840) and GG (P=0.867, OR=0.911, CI=95%; 0.308-2.699) had no significant associations with risk of breast cancer.
Conclusion: There was no significant association between FokI polymorphisms of the XRCC3 and risk of susceptibility to breast cancer, which was in accordance to some researchers. FokI polymorphisms of XRCC3 gene cannot be used as a biomarker in clinical predictive studies in relation to risk of breast cancer.
Methods: This case-control study was carried out on the women with breast cancer and healthy women located in Markazi province at Arak University Research, Iran, from October 2016 to March 2017. In the present study, the association of FokI polymorphisms and the risk of breast cancer was assessed by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques. In this method, genomic DNA was extracted from blood samples using the kit procedure. Then, PCR was performed and the SNP-containing DNA amplicons were subjected to digestion of enzymes. Following digestion, each sample was immediately analyzed by 3% agarose gel electrophoresis. Statistical analysis was done using SPSS and SNP Analyzer softwares and the final results were determined.
Results: No statistically significant difference was observed between the two groups of patients and controls for three genotypes the site rs1799794 (P=0.435). Genotype AG (P=0.384, OR=0.614, CI=95%, 0.205-1.840) and GG (P=0.867, OR=0.911, CI=95%; 0.308-2.699) had no significant associations with risk of breast cancer.
Conclusion: There was no significant association between FokI polymorphisms of the XRCC3 and risk of susceptibility to breast cancer, which was in accordance to some researchers. FokI polymorphisms of XRCC3 gene cannot be used as a biomarker in clinical predictive studies in relation to risk of breast cancer.
Mehrdad Mohammadi, Jamshid Faghri,
Volume 77, Issue 4 (7-2019)
Volume 77, Issue 4 (7-2019)
Abstract
Background: Staphylococcus aureus is a common pathogen in human that can be the cause of a wide range of infectious diseases including bacteremia, pneumonia, cellulitis, and osteomyelitis and skin and soft tissue infections. The coagulase enzyme is one of the most important virulence factors of this bacterium. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) Coa pattern is one of the molecular base typing methods. Molecular typing plays an important role in epidemiological studies of nosocomial infection, such as methicillin-resistant Staphylococcus aureus (MRSA) infection. The PCR-RFLP Coa gene technique provides a useful preliminary method to monitor variations in MRSA populations. We were done Coa-RFLP typing according to the method of Hookey et al., with some modifications.
Methods: In this cross-sectional study, one-hundred fifty isolates of S. aureus from urine and blood samples of patients that collected from educational hospitals of Imam Hossein and Al Zahra Isfahan University of Medical Sciences, Iran, from February 2018 to October 2018 were analyzed. After bacterial confirmation of isolates by Coa gene in polymerase chain reaction (PCR) technique, to perform coagulase gene typing, the repeated units encoding hypervariable regions of the coagulase gene of S. aureus were amplified by PCR. This was followed by AluI restriction enzyme digestion and analysis of restriction fragment length polymorphism (RFLP) patterns.
Results: Of 150 samples, 45 isolated of S. aureus were confirmed by biochemical methods. Of previous positive samples, 36 (80%) isolates carried Coa gene. Two different genotypes of Coa gene were obtained that include bp680 fragment in 20 specimens and bp750 fragment in 16 specimens. After enzymatic digestion by AluI restriction enzyme for RFLP, four different restriction patterns were obtained that including, the 280+400 pattern in 16 specimens (44.4%), 280+470 pattern in 7 specimens (19.4%), 340+340 pattern in 6 specimens (16.6%) and 750 patterns without digestion were in 7 specimens (19.6%).
Conclusion: Using the present experiments, it was determined that the PCR-RFLP pattern, 280+400, was the dominant pattern in the Staphylococcus aureus samples isolated in Isfahan.
Methods: In this cross-sectional study, one-hundred fifty isolates of S. aureus from urine and blood samples of patients that collected from educational hospitals of Imam Hossein and Al Zahra Isfahan University of Medical Sciences, Iran, from February 2018 to October 2018 were analyzed. After bacterial confirmation of isolates by Coa gene in polymerase chain reaction (PCR) technique, to perform coagulase gene typing, the repeated units encoding hypervariable regions of the coagulase gene of S. aureus were amplified by PCR. This was followed by AluI restriction enzyme digestion and analysis of restriction fragment length polymorphism (RFLP) patterns.
Results: Of 150 samples, 45 isolated of S. aureus were confirmed by biochemical methods. Of previous positive samples, 36 (80%) isolates carried Coa gene. Two different genotypes of Coa gene were obtained that include bp680 fragment in 20 specimens and bp750 fragment in 16 specimens. After enzymatic digestion by AluI restriction enzyme for RFLP, four different restriction patterns were obtained that including, the 280+400 pattern in 16 specimens (44.4%), 280+470 pattern in 7 specimens (19.4%), 340+340 pattern in 6 specimens (16.6%) and 750 patterns without digestion were in 7 specimens (19.6%).
Conclusion: Using the present experiments, it was determined that the PCR-RFLP pattern, 280+400, was the dominant pattern in the Staphylococcus aureus samples isolated in Isfahan.
Zahra Fasihizade , Bahram Ahmadi , Gholam Reza Shokoohi , Nilufar Jalalizand , Marjan Motamedi , Hossein Mirhendi ,
Volume 77, Issue 4 (7-2019)
Volume 77, Issue 4 (7-2019)
Abstract
Background: Dermatophytes create the most common fungal disease in humans, called dermatophytosis. The two species of Trichophyton rubrum and Trichophyton interdigital are responsible for over 80% of types of dermatophytosis. So far, several morphological and physiological methods have been used to differentiate these very similar species, but these methods are generally time-consuming and have low specificity. The purpose of this study was to introduce a simple and rapid duplex polymerase chain reaction (PCR) reaction to differentiate these two species from each other.
Methods: This research was an analytical and experimental study that was carried out from 2017 to 2018 in the Medical Mycology Laboratory, School of Public Health, Tehran University of Medical Sciences, Iran. For this purpose, the nucleotide sequences of the 4 regions of internal transcribed spacer (ITS), beta-tubulin, elongation factor 1 alpha and calmodulin in the two considered species of fungi were conducted bioinformatics analysis. The differences and similarities of nucleotides between two species in each of these genes were studied for selecting the primer. The specificity of selected primers was tested for duplex PCR reaction against sequenced isolates of dermatophyte species.
Results: According to the total data, the specific primers were selected from elongation factor 1 alpha gene. These primers produced a product of 173 and 384 bp, in Trichophyton rubrum and Trichophyton interdigital, respectively. They had high specificity in the face of various dermatophytes. The length of nucleotide sequences found in the genebank of this gene in the two species is between 700 and 770 bp. The similarity of the two species in this region is 94.6% and differs by 78 bp. Of the 107 extracted DNAs from clinical dermatophyte isolates, in duplex PCR 24 isolates were positive with Trichophyton interdigital primer and 71 isolates against Trichophyton rubrum. The remaining isolates, which included 6, were negative in this reaction, which included other dermatophyte species.
Conclusion: This method is a specific and fast differential method compared to conventional methods for identifying Trichophyton rubrum and Trichophyton interdigital from each other.
Methods: This research was an analytical and experimental study that was carried out from 2017 to 2018 in the Medical Mycology Laboratory, School of Public Health, Tehran University of Medical Sciences, Iran. For this purpose, the nucleotide sequences of the 4 regions of internal transcribed spacer (ITS), beta-tubulin, elongation factor 1 alpha and calmodulin in the two considered species of fungi were conducted bioinformatics analysis. The differences and similarities of nucleotides between two species in each of these genes were studied for selecting the primer. The specificity of selected primers was tested for duplex PCR reaction against sequenced isolates of dermatophyte species.
Results: According to the total data, the specific primers were selected from elongation factor 1 alpha gene. These primers produced a product of 173 and 384 bp, in Trichophyton rubrum and Trichophyton interdigital, respectively. They had high specificity in the face of various dermatophytes. The length of nucleotide sequences found in the genebank of this gene in the two species is between 700 and 770 bp. The similarity of the two species in this region is 94.6% and differs by 78 bp. Of the 107 extracted DNAs from clinical dermatophyte isolates, in duplex PCR 24 isolates were positive with Trichophyton interdigital primer and 71 isolates against Trichophyton rubrum. The remaining isolates, which included 6, were negative in this reaction, which included other dermatophyte species.
Conclusion: This method is a specific and fast differential method compared to conventional methods for identifying Trichophyton rubrum and Trichophyton interdigital from each other.
Related Websites
Site Keywords
Cardiology, Anatomy, Biology, Epidemiology, Immunology, Neuroscience, Physical therapy, Physiology, Dermatology, Endocrinology and metabolicSite Statistics
- Registered users: 7811 users
- Online users: 0 users
- Guest users: 107 users
- All visits: 16489784 visits
- Visits in 24 Hours: 11516 visits
- Total articles: 11373 articles
- Published articles: 4964 articles
© 2025 , Tehran University of Medical Sciences, CC BY-NC 4.0
Designed & Developed by : Yektaweb