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Showing 48 results for Polymerase Chain Reaction

Mohammad Mehdi Soltan Dallal, Shirin Nezamabadi, Jalal Mardaneh, Zahra Rajabi, Abolfazl Sirdani,
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)
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.
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)
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.
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)
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.

Hoda Ahmadi , Reza Mirfakhraie , Shiva Irani ,
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.

Sajad Shafai , Elham Moslemi , Mehdi Mohammadi , Kasra Esfahani , Amir Izadi ,
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.

Nahid Arefi Lisar , Parivash Kordbacheh , Sasan Rezaie , Mahin Safara , Roshanak Daie Ghazvini , Heidar Bakhshi , Zahra Omidvar Jalali ,
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.

Seyedeh Hakimeh Rezazadeh, Reza Shirkoohi, Abdolhamid Angaji, Seyed Yusef Seyedena, Amir Nader Emami Razavi,
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.

Leila Hassani , Golnaz Asaadi Tehrani , Sina Mirza Ahmadi ,
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.

Milad Pezeshki , Jamshid Ansari ,
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.

Mehrdad Mohammadi, Jamshid Faghri,
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.

Zahra Fasihizade , Bahram Ahmadi , Gholam Reza Shokoohi , Nilufar Jalalizand , Marjan Motamedi , Hossein Mirhendi ,
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.

Marzieh Kazerani , Nahid Jalalian Elahi , Najmeh Mohajeri , Kiarash Ghazvini , Sara Taghdisi , Mohmadreza Ghafghazi , Mahdieh Motaghi , Mahdieh Motaghi ,
Volume 77, Issue 7 (10-2019)
Abstract

Background: Molecular detection has recently been proposed by nucleic acid amplification, known as polymerase chain reaction (PCR). The aim of this study was to compare the diagnostic method of smear and polymerase chain reaction with culture in terms of sensitivity, specificity, positive and negative predictive value in the diagnosis of pulmonary tuberculosis.
Methods: In this cross-sectional study, sputum samples were collected from 58 patients with suspected pulmonary tuberculosis referred to Ghaem Hospital in Mashhad from the beginning of April 2017 to the end of March 2018. The samples were delivered to the laboratory in less than 72 hours. Patients were sampled for three times. Bronchoscopy and Broncho alveolar lavage were performed in patients who were unable to produce sputum. The smear test was reported by Ghaem’s Laboratory after 24 hours. In our study, the culture method was considered as the gold standard and the sensitivity and specificity of the PCR methods and smear were compared with it.
Results: Patients ranged in age from 18 to 89 years. Among 58 suspected pulmonary tuberculosis, the method of cultivation confirmed the presence of the disease in 25 cases (43.1%). However, with smear, the presence of the disease has been proved in 27 patients (46.6%) and with the method of PCR in 24 patients was (41.4%). Sensitivity of smear in the diagnosis of pulmonary tuberculosis was (100%), and its specificity was 93.9%, the positive predictive value of this test was (92.6%) and the negative predictive value was (100.0%). The sensitivity of the PCR method in diagnosis of pulmonary tuberculosis was 88.0% and its specificity was 93.9%. The positive predictive value of this was (91.7%) and the negative predictive value was (91.2%).
Conclusion: In this study, between the two methods of smear and polymerase chain reaction, the acid fast smear method was more sensitive to the diagnosis of pulmonary tuberculosis than the polymerase chain reaction and the specificity of both methods were the same.

Masoomeh Babaei , Mehrdad Hashemi , Behzad Banieghbal ,
Volume 77, Issue 10 (1-2020)
Abstract

Background: Micro-Ribonocellic Acids (miRNA) are non-coding nucleic acids that are evolutionally protected and have a length of 24-20 nucleotides. MiRNAs control the expression of genes after transcription by mRNA degradation or translation inhibition. By blocking the oncogenic miRNAs and creating the necessary and functional miRNAs (tumor suppressor), these small regulatory RNAs can have therapeutic applications in cancer. The high mortality from lung cancer highlights the fact that the majority of patients are diagnosed at an advanced stage of the disease. The use of serum biomarkers can help early detection. MiRNA is more stable than mRNA. MiRNA expression in tissue, plasma, sputum, and urine samples can be detected by fixed formulation. In addition, miRNAs are important modulators of gene expression, diagnostic markers, and prognosis. Therefore, in the present study, the expression of miR-137 in the serum of patients with lung cancer was investigated.
Methods: In this descriptive and analytical study, 100 serum samples were collected from patients referring to Masih Daneshvari Hospital in Tehran from August 2017 to May 2018. Also, individual and clinical information were collected by a questionnaire and real-time polymerase chain reaction (RT-PCR) was used for the qualitative evaluation of changes in expression of miR-137.
Results: Data showed that there was no significant difference between the expression of miR-137 in serum samples of the first and second stages of the disease. While in the serum of patients with lung cancer who metastasized in the third and fourth stages, miR-137 expression decreased by 3.2 (P=0.42) and 6.8 times (P=0.003), respectively. Based on the results, it can be inferred that the measurement of miR-137 expression in lung cancer patients with concomitant reduction can be a sign of the progression of the disease.
Conclusion: Based on the results of this study, there was a significant relationship between miR-137 expression and lung cancer.

Azam Shiralinezhad , Farzaneh Firoozeh , Mansooreh Momen Heravi, Esmat Aghadavod , Mojtaba Sehat ,
Volume 77, Issue 10 (1-2020)
Abstract

Background: Sepsis or blood stream infection is a clinical lethal syndrome with severe systemic inflammatory response to infection, if not treated quickly, is associated with dangerous consequences and high morbidity and mortality. The traditional and conventional method for identification of sepsis is blood culture method which is so time-consuming and long that it eliminates the possibility of rapid treatment. Although, new molecular methods, due to their high sensitivity, specificity, and speed, lead to the rapid and accurate and exact detection of bacterial sepsis within only a few hours. The aim of this study was diagnosis of bacteremia in patients with suspected sepsis using amplification of 23S rRNA gene by polymerase chain reaction (PCR).
Methods: This cross-sectional study was performed in two clinical and analytical steps at Shahid Beheshti University Hospital in Kashan City, Iran, in twelve months from November 2016 to December 2017. The blood samples of two hundred and fifty-six patients with suspected sepsis admitted to Shahid Beheshti Hospital were studied by PCR method using specific primers of 23S rRNA gene of the bacteria.
Results: The finding of molecular assays using PCR showed that of 256 blood samples that were collected from patients with clinical signs and symptoms of sepsis, 80 (30.2%) diagnosed with bacteremia. Of these patients diagnosed with sepsis, 46 out of 80 (57.5%) were male while 34 out of 80 (42.5%) were female. The most PCR positive results were obtained among patients with diabetes and bedsore as underlying diseases (21.3%). Statistical analysis showed that there was a significant correlation between results of molecular methods by PCR assays and history of antibiotic use. 
Conclusion: Overall, the results of the present study showed that the molecular methods such as polymerase chain reaction using universal 23S rRNA primers is an appropriated test for diagnosis of bacteremia in blood samples of patients with suspected sepsis.

Majid Gholipour , Mastaneh Seifabadi , Mohammad Reza Asad ,
Volume 77, Issue 11 (2-2020)
Abstract

Background: Skeletal muscle mass, which is regulated by a balance between muscle protein synthesis and degradation, is an important factor for movement to meet everyday needs, especially in pathological conditions and aging. The purpose of the present investigation was to compare the alterations of the gene expression involved in muscle protein synthesis and degradation signaling pathways induced by two exercise training protocols.
Methods: Eight weeks old Wistar rats have been assigned to the present experimental study, which was conducted from August 2018 to October 2018 at the animal laboratory of Tehran University. They were randomly divided into two resistance and endurance training groups and one control group, and run on a treadmill, 5 sessions per week for 8 weeks. 48 hours after the last exercise session, the rats in the two groups were anesthetized, and the dissected soleus muscles from euthanized animals were stored at -80° for RT-PCR and Western blot analysis later. Between-group differences were analyzed by the parametric and non-parametric tests for normally and non-normally distributed data respectively, at the significance level of α˂0.05.
Results: Compared with the control group, mTORC1 gene expression was increased significantly just in the endurance group (P=0.022), whereas both endurance and resistance exercise protocols caused a significant increase in Rps6kb1 (P˂0.001 and P=0.001 respectively). In protein degradation pathway, although, FOXO3a did not alter significantly (P=0.463), eIF4Ebp1 gene expression was inhibited by both endurance and resistance exercise training protocols (P˂0.001 and P=0.001 respectively). The alterations of Rps6kb1 and FOXO3a gene expression were confirmed by Western blot analysis.
Conclusion: The results showed that the exercise training protocols of the present study had approximately similar effects on alterations of gene expression involved in skeletal muscle protein synthesis and degradation pathways. Therefore, application of the protocols may be considered to prevent or reduce the muscle atrophy in pathological conditions such as motor neuron disease, aging, and/or muscle strength improvement in athletes.

Amin Derakhshanfar , Hadi Tavakkoli , Javad Moayedi , Ali Poostforoosh Fard ,
Volume 77, Issue 12 (3-2020)
Abstract

Background: The H9N2 subtype of the influenza virus, which is endemic in many regions of Iran, is considered as a candidate for future pandemics. In the present study, excretion time of the Iranian endemic influenza virus (H9N2 subtype) from the feces and pharyngeal secretions of laying chicken breeds was evaluated.
Methods: This experimental study conducted at the Diagnostic Laboratory Sciences and Technology Research Center of Shiraz University of Medical Sciences, and the Department of Clinical Science in School of Veterinary Medicine of Shahid Bahonar University of Kerman, from June 2017 to September 2017. At first, the influenza virus A/Chicken/Iran/SH-110/99 (H9N2) was cultured in the allantoic fluid of the embryonated egg and the EID50 for virus was determined by Reed and Muench method. Afterward, the Hy-Line chicks were inoculated intranasally with 106 EID50/ml of influenza virus (H9N2 subtype) and samples were collected from the oropharynx and feces of the birds on days 2, 5, 10 and 17 after inoculation. The presence of the virus in the samples of challenged birds was assessed using the real-time polymerase chain reaction (PCR) method.
Results: The influenza virus was shed from the oro-pharyngeal secretion and feces of the birds 2 days post-infection, and continued until days 10 and 17, respectively. In comparison to the oro-pharynx, the virus was recovered in the feces for a longer time. The influenza virus was detected in 100% and 57.1% of oro-pharyngeal and feces samples of the infected birds on day 2, 85.7% and 100% on day 5, 28.6% and 71.4% on day 10, and 0% and 28.6% on day 17 post-inoculation, respectively. The maximum risk of infected chicken for humans is seen from 2 to 5 days post-infection.
Conclusion: Detection of virus in the samples of birds that challenged with the H9N2 influenza virus showed that the virus could shed from the feces to the surrounding environment longer than the pharyngeal secretions and could be hazardous to humans in contact.

Zeinab Borjian Boroujeni , Mohammad Kord, Zeinab Tabanejad, Seyed Saeed Asadi, Morteza Mesri , Mohammad Panji, Mahdi Zareei,
Volume 78, Issue 12 (3-2021)
Abstract

Background: Trichosporon species are commonly known as causative agents of skin infections and also responsive in some other systemic and disseminated diseases, especially in immunocompromised patients and those with leukemia or lymphoma. Chronic cutaneous infections with Trichosporon have been reported in non-immunocompromised patients, too.
Case Presentation: This study is a case report of tinea pedis caused by Trichosporon asahii in an immunocompetent 39-year-old man who was a member of the military force with continuous wearing of army boots during his daytime work. In April of 2019, after visiting a dermatologist, he was referred to the Ghaem medical mycology laboratory of the Department of Health, Rescue and Treatment of Iran Police Force in Tehran. Clinical symptoms were scaling and erythematous patches on his left foot with intensive itching for four-months. In the laboratory, macroscopic and microscopic examination using direct 15% KOH wet mount was carried out as well as culture methods on fungal media (Sabouraud's dextrose agar with and without cycloheximide and chloramphenicol). According to microscopic observation and appearance of culture media colonies, the diagnosis was Trichosporon genus as the fungal agent of disorder. Molecular method analysis (PCR) using amplification of ITS region with universal primers (ITS1 and ITS4) and sequencing identified Trichosporon asahii as a causative species of the disease. The patient was treated with topical clotrimazole (twice/day) and oral fluconazole (150 mg/day) for four weeks, and recovered.
Conclusion: The conditions that cause the feet to be exposed to excessive sweating, moisture and darkness for a long time (wearing shoes and army boots for a long time); increase the risk of skin fungal infection. Therefore, full respect for an individual healthy manner seems to be essential.

Jamshid Ansari, Milad Pezeshki, Azam Ahmadi, Ali Chehrei,
Volume 81, Issue 9 (12-2023)
Abstract

Background: Lung cancer has the highest incidence and mortality rate of all cancers worldwide. In Iran, it is one of the commonly diagnosed malignancies, and its frequency is increasing rapidly. Genetic variants in DNA repair genes are linked to differences in efficiency of repairing DNA damage, which can influence lung cancer susceptibility. EXO1 is a key gene involved in the mismatch repair pathway. The K589E polymorphism in EXO1 may alter the DNA repair activity of the encoded protein and impact lung cancer risk. The aim of this study was to investigate associations between the K589E polymorphism in EXO1 and lung cancer risk in the Iranian population, and evaluate its potential as a prognostic biomarker.
Methods: This case-control study was conducted to investigate EXO1 K589E variant with susceptibility to lung malignancy in the Iranian population. One hundred patients with lung cancer as a patient group and 100 healthy individuals from Khansari Hospital located in Markazi province were studied, from January 2020 to May 2022. DNA extraction from blood samples of participants was done using a kit.  Genotype determination of both patient and control groups was done using PCR-RFLP technique. Finally, statistical results were analyzed using SPSS software and the logistic regression method.
Results: Genotype and allele frequency  analysis showed the AA genotype (P=0.004, OR=5.391, 95% CI: 1.690-17.200) and A allele (P=0.010, OR=2.851, 95% CI: 1.291-6.300) were correlated with susceptibility to lung cancer. On the other hand, people carrying the G variant allele had a lower risk of lung cancer.
Conclusion:  In summary, this study found the AA genotype and A allele of K589E in EXO1 are correlated with risk of lung cancer in Iranians, while the G allele has protective effects. The K589E polymorphism may serve as a prognostic biomarker for lung cancer susceptibility, but more studies with high population size are required.

Shokoufeh Akbari, Monireh Rahimkhani, Reza Mirnejad,
Volume 81, Issue 10 (1-2024)
Abstract

Background: Today, Methicillin-Resistant Staphylococcus Aureus (MRSA) has become one of the public health concerns due to its resistance to antimicrobial drugs, and this problem makes treating patients with infections caused by this bacterium difficult. Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) strains are pervasive in both community and hospital settings, primarily attributable to Staphylococcus aureus' capacity to colonize areas like the nose or skin. In this study, with the aim of comparing phenotypic (disc diffusion method) and genotypic (PCR) methods, to detect methicillin-resistant Staphylococcus aureus isolated from patients of hospitals under supervision of Tehran university of medical sciences, and also detection of nor A, that is the one of the most important genes in efflux pump cluster genes.
Methods: The present research was a cross- sectional study that was conducted from February 2022 to September 2023. In this research, 43 isolated strains of Staphylococcus aureus from wound discharge and blood samples, were collected from different departments of Tehran hospitals and had submitted to the research laboratory of the school of allied medical sciences in Tehran university of medical sciences. After identifying the strains, the resistance of the isolates to 14 types of antibiotics was checked by disk diffusion method.
Results: Staphylococcus aureus diagnostic tests including gram staining on colonies, catalase, coagulase, DNase tests were performed and it was found that all strains were Staphylococcus aureus. In the next step, all samples were resistant to Cloxacillin by disc diffusion method, and the presence of mec A gene in them was confirmed by PCR method, thus the presence of MRSA strains was confirmed from the genotypic point of view. Of the 43 Staphylococcus aureus strains, 26 samples were identified as having the nor A gene by PCR and electrophoresis.
Conclusion: The results of the present research have shown that the prevalence of Staphylococcus aureus bacteria in hospital samples is significant and resistance to methicillin and ciprofloxacin has increased in the strains of this bacteria.


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