Tehran University Medical Journal

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Macrophage colony stimulating factor (M-CSF) has previously been shown to affect the differentiation of cells of the mono-nuclear phagocytic line. More recent studies indicate that M-CSF may have a role in pregnancy. In the present study, the expression of M-CSF in the human placenta was demonstrated. Placental mRNA was isolated and used as template for synthesis of complementary DNA (cDNA). The presence of M-CSF related sequences in the cDNA was shown by PCR and RT-PCR reactions in which M-CSF specific primers were used. In addition, it was shown that a 2.4 kb cDNA after electrophoresis and transfer to a nylon filter, hybridized with a digoxygenin labeled M-CSF specific probe.

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Background: Staphylococci as a micro-organism, has the most importance to cause nosocomial infections, particularly in patients with indwelling catheters or other medical devices. Unfortunately 90% of Staphylococci isolated from the nosocomial infections are resistant to methicillin, and methicillin resistance strains are also resistant to a wide range of antimicrobial drugs, therefore detecting of these strains are valuable to eradicate the infection elements. Despite guidelines published by the national committee for clinical laboratory standards (NCCLS) for testing of susceptibility to methicillin for Staphylococci, the phenotypic method for detecting methicillin resistance remains controversial. Therefore, the genetic assays have been used to detect antimicrobial susceptibility of Staphylococci to methicillin.
Materials and Methods: Resistance to methicillin is coded by mec A gene in staphylococcus, and this gen must be detected in genetic assays. In this study 155 clinical staphylococcal isolates (70 coagulase- negative staphylococcus and 85 coagulase- Positive staphylococci) were evaluated for susceptibility to methicillin by using disk diffusion method.
ResuIts&Conclusion: Methicillin resistance was shown in 62 coagulase- negative staphylococcus (72.9%) and 27 coagulas positive staphylococcus (38.6%) but 63 coagulase negative Staphylococci (74%) and 28 coagulase positive isolates with mec a gene associated resistance were detected by PCR method. The results of this test were compared to the results for mec A gene detection by PCR test as a gold standard. The sensitivity, specifity and accuracy of the disk diffusion test for coagulase-negative staphylococcus were 96.8%, 95.45% and 96.47% and for coagulase positive staphylococci were 98.43%, 95.45% and 98.32% respectively.

 

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Background: Cervical cancer is the second leading cause of cancer death among women. In this cancer, the effects of prevention, early diagnosis and treatment more than other cancers decrease the mortality rate. In 1970 human papilloma virus (HPV) was introduction as major etiologic factor of cervical cancer. Different studies throughout the world revealed strong correlation between HPV and cancerous & precancerous changes in epithelial cells. Since cell culture and serological methods can not recognize the virus and its subtypes, the importance of the molecular methods including polymerase chain reaction (PCR) in early and definite diagnosis of virus is obvious.

Methods: In this study, after patient selection using the related protocol and completion of the questionnaires, 100 samples from cancer lesions of cervix selected. Then DNA extraction from paraffin blocks performed using standard method. Multiplex PCR with two pairs of primer (one as internal control) performed and the PCR product run on 8% polyacrylamid gel.

Results: The results showed that 73% of the tissues were infected by HPV.

Conclusion: This finding confirm the previous results based of correlation between HPV,and cervical cancer.

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Background: Tuberculosis is still one of the most important causes of mortality and morbidity in many countries and is the second only to human immunodeficiency virus as a cause of death worldwide resulting from a single infectious agent. In 1993, the World Health Organization declared tuberculosis a global public health emergency. Conven-tional methods for the diagnosis of Mycobacterium tuberculosis (MTB) infections are time consuming, as MTB culture requires 3-8 weeks for growth. To determine the sensitivity of polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), we have evaluated Mycobacterium tuberculosis DNA in peripheral blood samples with PCR technique in adults with new cases of pulmonary and extra-pulmonary tuberculosis. Setting: Department of Infectious disease of Imam Khomeini Hospital, 2004- 2005, Tehran, Iran.

Methods: In this cross-sectional study, we evaluated MTB DNA extracted from 3ml citrated peripheral blood samples from 95 adults with new cases of pulmonary and extra-pulmonary tuberculosis. DNA extraction was performed using a commercial PCR kit with IS1081 primers. For prevention of cross contamination and reduction of false positives, all steps were performed under laminar hood.

Results: The 95 patients, 59 of whom were male, had a mean age 44.44 years (SD±20.26) 69 cases had pulmonary and 26 had extra-pulmonary tuberculosis. PCR was positive in 32 (33.7%) patients and negative in 63 (66.3%) cases. The overall sensitivity and accuracy of the PCR assay was 44.1% for pulmonary, 19.2% for extra-pulmonary and 10% for disseminated tuberculosis, respectively.

Conclusion: The low sensitivity of the IS1081 primer MTB-PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis. However, further studies are needed to confirm this technique as an alternative test for the diagnosis of tuberculosis.

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Background: Cervical cancer is the second most common cancer of the women worldwide. It is also an important cause of cancer-related mortality in women, after breast cancer. Nearly half million of new cases are identified yearly. The incidence rate in developing countries is greater than the developed countries. Epidemiologic studies have shown that the association of genital human papilloma virus (HPV) with cervical cancer is strong, independent of other risk factors, and consistent in several countries. The aim of this study was to determine the frequency of HPV in patients with high grade cervical intraepithelial neoplasia (CINIII, CIN II) and squamous cell carcinoma (SCC) of cervix.
Methods: Hundred specimens from patients with SCC and CINIII, CIN II, confirmed by histological review, referring to Mirza Koochak Khan Hospital from 1999-2004 were enrolled in a cross sectional study. Polymerase chain reaction was utilized for identification and typing of HPV DNA. To increase the sensitivity of HPV detection, nested PCRs were performed using MY09/MY11 as outer and GP5/GP6 as inner primers.
Results: It was possible to extract 77 of 100 specimens that HPV DNA was detected in 47 of 77 specimens. Infection with HPV was present in 32 specimens (86.5%) among SCC patients and in 15 specimens (37.5%) among CINIII, CIN II patients. The most frequent HPV types in SCC patients were HPV 16 and 18 (59.38%) and then 33 (34.38%) and in CINIII, CIN II patients was 16 (53.33%) and 18 (40%). the most frequent co-infection in both groups was HPV 16 and 18 which was present in 40.62% and 26.7% of cases respectively.
Conclusions: The most frequent HPV types in patients with SCC and CINIII, CIN II was 16 and 18 that is identical to many other countries infection pattern.

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Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: Vulvovaginal candidiasis is a fungal disease with itching, and vaginal thick white discharge. Most of non-albicans species have less sensitivity to azoles. So, definition of candida species which lead to vulvovaginal candidiasis is very important to perfect usage of drugs. In the present study 191 Candida isolates from 175 patients who admitted in Gynecology department of Mahdieh Hospital during the period 1385-1387 were identified by multiplex PCR.
Methods: One hundred seventy five vaginal swab specimens from patients were cultured on Sabouraud Dextrose Agar (SDA). The internal transcribed spacer 1 (ITS1) region between the 18S and 5.8S rRNA genes and a specific DNA fragment within the ITS2 region of Candida albicans were amplified and the multiplex PCR products were separated by electrophoresis in 2% agarose gel (200 mA, 140V), visualized by staining with ethidium bromide, and photographed.
Results: One hundred ninety one Candida isolates were identified in vaginal swab specimens from 175 patients. In 89.7% of cases, single candida species and in 10.3% cases, multiple candida species were isolated. C. albicans (65.1%), C. glabrata (13.1%), C. tropicalis (6.2%), C. krusei (4%), C. guilliermondii (0.6%), C. parapsilosis (0.6%), C. glabrata and C. albicans (5.7%), C. albicans and C. parapsilosis (1.1%), C. glabrata and C. tropicalis (0.6%), C. krusei and C. tropicalis (0.6%), C. albicans and C. tropicalis (0.6%), C. krusei and C. albicans (0.6%), C. glabrata and C. krusei (0.6%), and C. glabrata and C. krusei and C. albicans (0.6%) were the cause of disease.
Conclusion: Our findings suggest that, the common cause of both recurrent and non-recurrent vulvovaginal candidiasis was C. albicans, and then C. glabrata. Also the most common mixtures of Candida species were combination of them

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800x600 Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background: In the last two decades, cryptococcosis has been gaining a distinct public health importance due to the growing number of AIDS cases. Considering the low sensitivity of direct examination with India ink and culture, use of sensitive techniques is crucial in the diagnosis of cryptococcal meningitis. Polymerase Chain Reaction (PCR) can be used to directly detect Cryptococcus neoformans in CSF samples to increase the diagnostic power in cases where conventional methods are unable to detect the organism.
Methods : In this cross-sectional study, CSF samples were obtained from 25 patients suspected of having neurocryptococcosis. The patients were referred to the Medical Mycology Laboratory of the School of Public Health affiliated to Tehran University of Medical Sciences from March 2009 to February 2010. Three different methods, direct India ink examination, culture and PCR were used to evaluate the CSF samples. Two 102 and 106 of Cryptococcus neoformans dilutions in 1ml of CSF were prepared and examined by the three methods. In PCR method, two primer pairs were selected to amplify the Cryptococcus neoformans URA5 gene. The sequences of primers were for A, B, C and D serotypes.
Results : Only in one case PCR, as well as direct examination and culture were positive. All the other samples were negative in PCR, direct examination or culture. Both CSF dilutions were positive in the three tests in the mentioned patient and the positive control.
Conclusion: PCR method can efficiently identify both control and positive samples of Cryptococcus neoformans.

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Background: In the Mediterranean region , Kaposi's sarcoma (KS) has a high prevalence especially in patients with AIDS. Iran is located close to the Mediterranean region and the HIV prevalence is increasing in our country . In some stages, Kaposi's sarcoma is morphologically similar to other vascular tumors. Owing to the presence of human herpesvirus 8 (HHV-8) in all cases of Kaposi's sarcoma , detection of virus DNA by PCR method can help in the identification of non-diagnostic cases. Moreover, the prevalence of HHV-8 genotypes is different in various regions of the world and in different races. There are limited studies performed on the HHV-8 genotypes in Iranian population.

Methods: Patients with Kaposi's sarcoma from 2001 to 2011 who refer to Tehran Razi Hospital were enrolled in this study. HHV-8 DNA was extracted from paraffin blocks and amplification of the virus genome was performed by PCR method . Finally, the target DNA fragment was used for sequencing and genotype determination.

Results: PCR was performed on 53 cases. In 8 cases with suspicious morphology, PCR was negative and they were excluded from study. Of remaining 45 cases, 35 had positive PCR results, 7 had negative results and 3 had low PCR product. Samples from 28 cases that had positive PCR results, which were acceptable for genotyping, were chosen for sequencing. Twenty cases had genotype C, 7 cases had genotype A and one case was negative. The results are consistent with other studies in our geographical area. No correlation was found between the different microscopic stages and HHV-8 Genotypes.

Conclusion: Since the HHV-8 is obtained in almost 100% of KS lesions and PCR s ensitivity in detection of the virus is close to 100 %, KS diagnosis can be confirmed in suspicious cases by detection of HHV-8 DNA on paraffin blocks. Moreover the prevalence of HHV-8 genotype was determined in Iran.

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Background: The aim of this study was compared the efficacy of  the designed primers and already published primers for detection of the exoA, oprL and algD genes by PCR assay  for finding a rapid, accurate and highly sensitive and specific procedure to detect the Pseudomonas aeruginosa in the serious and fatal infections such as cystic fibrosis disease, burned individual.
Methods: A total of 150 clinical specimens were inoculated in to routine and selective culture media for Pseudomonas aeruginosa isolation. Specific primers were designed by bioinformatics analysis for detection of the virulence genes exoA, oprL and algD. The available sequences of these three genes were obtained from NCBI and multiple alignments were performed to find the conserved sequences of each gene for primer designing. Both multiple alignment and primer designing steps were carried out by AlleleID software, version 7.0.
Results: Microbiological culture methods were showed that 70 Pseudomonas aeruginosa strains isolated from the 150 clinical specimens. PCR assay performed by using the designed primers shown 68, 70 and 69 positive results from 70 direct specimens for exoA, oprL and algD respectively that shown 97.2%, 100% and 98.6% sensitivity for above genes. PCR assay performed by using the already published primers shown 57, 49 and 28 positive results for above genes respectively that shown 81.5%, 70% and 40% sensitivity.
Conclusion: The present study shows that by using the high specific primers for detection of the mentioned genes of the Pseudomonas aeruginosa. The conventional PCR assay detected the early colonization of the organism in Cystic Fibrosis patients with more sensitivity and specificity before several mounts to obtain positive culture. Indeed PCR assay with high specific primers has more sensitivity and specificity as a rapid and accurate diagnosis of the organism in other deadly infections by using the direct clinical specimens.

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Background: Colorectal Cancer (CRC) is the third common cancer in the world. One of the pathways in colorectal tumor genesis is Microsatellite Instability (MSI+). MSI is detected in about 15% of all colorectal cancers. Colorectal tumors with MSI have dis-tinctive features compared with Microsatellite Stable (MSS) tumors. Due to the high percentage of MSI+ in patients with CRC in Iran, screening of this type of CRC is im-perative. In current study, two markers (BAT-26 and BAT-25) were used to determine an appropriate screening technique with high sensitivity and specificity to diagnose MSI status in patients with CRC. Methods: Allelic variation in two markers (BAT-26 and BAT-25) was analyzed in tis-sues and sera of 44 normal volunteers and tumor and matched normal mucosal tissues as well as sera of 44 patients with sporadic colorectal cancer by Real Time PCR (Hy-bridization probe) and High-Performance Liquid Chromatography (HPLC) techniques. The sensitivity and specificity of Real Time PCR and HPLC compared with sequencing as gold standard. The data were statistically analyzed using Student’s t-test and 2 or fisher exact test, where applicable with (P<0.05). Receiver-operating-characteristic (ROC) curves were used to evaluate the sensitivity and specificity. Results: The sensitivity and specificity of BAT-26 with Real Time PCR method (Hy-bridization probe) were 100% in comparison with gold standard method. Whereas the sensitivity and specificity of BAT-26 and BAT-25 with HPLC were 83%, 100% and 50%, 97%, respectively. Neither HPLC nor Real time PCR could detect circulating DNA with MSI property in sera. Conclusion: The sensitivity and specificity of real time PCR in MSI detection is the same as sequencing method and more than HPLC. BAT-26 marker is more sensitive than BAT-25 and MSI detection with Real time PCR could be considered as an accu-rate method to diagnose MSI in CRC tissues not sera.

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Background: Toxic shock syndrome (TSS), a dangerous consequence of Toxic shock syndrome toxin-1 (TSST-1) caused by Staphylococcus aureus. The early detection for infections of Staphylococcus aureus in burned children is very important, also the pre-vention for consequences of TSST-1. Fever is one of the most noticeable sign in burned children. On the other hand, fever is one of the important consequences of TSST-1 pro-duction. Methods: This study aimed to assess the toxic shock syndrome toxin-1 level in the wound’s specimens of two groups febrile and afebrile in the hospitalized burned chil-dren in Motahari hospital Tehran, Iran in the year 2013. In this case-control study, 90 children who admitted to the burn unit, divided in two groups of 45 patients: febrile (cases group) and afebrile (control group). All of burned children under went wound biopsy, and then all of wound’s specimens were tested by PCR for specific primer of toxin producing genome. Finally all of data collected and statistically analyzed. This data include group febrile and afebrile, demographic characteristics, percentage of burned surface severity and result of PCR. Results: The positive result for PCR test, production of TSST-1 in febrile burned chil-dren (cases group) was 37.7% and in afebrile burned children (control group) was 11.1% that this different was statistically significant (P=0.003). The mean and stan-dard deviation for percentage of burned surface (severity) in samples with positive re-sult for PCR test was 30.9±16.93 and in samples with negative result for PCR test was 20.09±11.02 that this different was statistically significant (P=0.01). There was no dif-ference between positive PCR result and negative PCR result of age and sex. Conclusion: Direct association was approved between the production of TSST-1 and the occurrence of fever in burned children. Increased surface severity of burns also re-lated to the production of TSST-1. Further research is recommended.

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Background: Pseudomonas aeruginosa is a gram-negative pathogens opportunism which causes severe infections in human beings. The most common infection include: endocarditis, meningitis, septicemia and chronic lung infections in cystic fibrosis pa-tients. This bacterium has many pathogenic factors including exotoxin A, lipopoly-sacharide, phospholipase C, pili, elastase and alkaline protease. The purpose of this study was to evaluate the frequency of exotoxin A gene (ETA) as a strong virulence factor and sensitivity determination of polymerase chain reaction (PCR) in pseudomonas aeruginosa isolated from second and third-degree burn patients. Methods: This study has performed in Besat University Hospital in Hamadan from January to December 2012. We used 170 isolated samples. The samples were isolated from blood and skin biopsy in second and third-degree burn patients. We had 79 strains positive culture of pseudomonas aeruginosa. Forward and reverse primers used for PCR were designed by DNASIS and Oligo software. Then genomic of known strains were extracted by DNA purification kit and indentified by PCR. The quality and quantity of the extracted DNA was determined using spectrophotometry. For determination of PCR sensitivity was used culture test as gold standard. DNA of pseudomonas aeruginosa (ATCC 27853) was used as a positive control. Finally data was analyzed using SPSS software. Results: Out of 170 isolated samples, 79 strains of pseudomonas aeruginosa isolated from burn patients had positive culture. PCR of isolated positive culture demonstrated that 5 strains (6.33%) were with out this virulence factor and 74 strains (93.67%) had ETA gene. So the sensitivity of test based on sensitivity formula was 94.04%. Conclusion: Our results showed that sensitivity of PCR mediated ETA gene in detection of pseudomonas aeruginosa strains is considerable and this factor can be used as a good factor identifying of pseudomonas aeruginosa. It seems more studies with larger sample size is necessary in this area.

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Background: Colorectal cancer is the third most common cancer in the world. Non-coding RNA especially miRNAs have important regulatory roles in cancer. miRNAs are small non coding RNA 21-23 nucleotides long which have different levels of expression between tumors and normal tissues. This study was designed to compare expression level of miRNA-21 between Iranian population colorectal cancer tissues and normal tissue. Methods: This case-control study has performed in medical genetics department of Tehran University of Medical Sciences from January to November 2013. We used 35 samples. The samples were isolated from tumor and adjacent normal tissues of colon. Thirty-five samples were divided into different groups according to cliniopathologic features including tumor size (>4 and <4 cm), metastasis (+ and -) and stage. After small RNA extraction from tissues by small RNA purification kit the quality and quan-tity of extracted RNA was determined using spectrophotometry. cDNAs were synthe-sized and real-time polymerase chain reaction carried out. Finally expression levels were statistically analyzed by LinRegPCR and REST software. Results: miRNA-21 expression ratio in stages I, II and III were 1/804 and 4/574, re-spectively, the increase from stage III was statistically significant (P= 0.037). The ex-pression were also studied according to different clinicopathologic status of colon can-cer, tumor size (>4 and <4 cm) and metastatic (+ and -), miRNA-21 over expressed in both groups, however the increase was not statistically significant. Conclusion: In this study, we found miR-21 over-expression in advanced stage in tu-moral tissue comparing with normal adjacent tissue. This means perhaps in the future it would be possible to use miRNA-21 as an informative prognostic biomarker to guide for better treatment strategies for colorectal cancer patients. Our findings also indicate that miRNA-21 is a promising new molecular target for designing novel therapeutic strategies to control colorectal cancer.

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Background: Mycoplasma contamination in cell cultures is considered as a major economic, research and production problem. In this study, mycoplasma-infected Vero cell lines were treated by various dilutions of ciprofloxacin and enrofloxacin in a timely manner. Removal of mycoplasma contamination from infected cell cultures was evaluated and demonstrated by polymerase chain reaction (PCR) method. Methods: This study was done from October 2013 to May 2014, in Human Rabies Vaccine Laboratory, Pasteur Institute Production and Research Complex, Tehran, Iran. Different dilutions of ciprofloxacin and enrofloxacin were used in sequential passages for treatment of infected Vero cell line. Based on lowest passages of the cell line, antibiotic treatment with ciprofloxacin and enrofloxacin was done. Amelioration of the infection and removal of mycoplasma contamination was confirmed in each step by PCR method. The technique for order of preference by similarity to ideal solution, TOPSIS method, was used to suggest the most efficient concentration of ciprofloxacin and enrofloxacin. Results: Proposed concentration of ciprofloxacin is 20 μg/ml, and in the second order is 200 μg/ml. For enrofloxacin the best proposed concentrations are 30, 300 and 3 μg/ml respectively. Ciprofloxacin and enrofloxacin and ability of them for removal of mycoplasma and also the time of treatment were verified by evaluation of the recurrence of infection through consecutive subcultures of the treated cell line. Conclusion: Our results showed that 20 μg/ml of ciprofloxacin was the dilution of choice for mycoplasma elimination followed by 200 μg/ml of ciprofloxacin. Concentrations of 3, 30 and 300 of enrofloxacin, respectively, are appropriate for mycoplasma removal. More detailed works would be needed to verify the authenticity of the proposed simple and affordable way of mycoplasma elimination.

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Background: Breast cancer is the second most common cancer in the world after lung cancer also is the fifth cause of cancer mortality. About 90 percent of cancer mortality is because of metastasis and devastating between cell attachments, especially tight cell junctions. Epithelial mesenchymal transition is a phenomena involved in metastasis and starts with cell detachment. Occludin is the integral membrane protein which is located in tight junctions. Obviously distressing tight junction, which facilitates the stages of metastasis in cancer cells are very critical step. The aim of this study was to demonstrate the importance of occludin expression and its relationship with invasiveness in human breast cancer. Methods: In a cross sectional study we evaluated 30 patients who were referred to Caner Institute of Imam Khomeini Hospital Complex, Tehran, Iran, from March 2013 to April 2013. Samples were derived from fresh frozen tumor of patients suffering from breast cancer after inform consent assignment in the Tumor Bank of Iran (TBI). RNA was extracted from tumor tissue followed by reverse transcription, polymerase chain reaction (PCR), conventional Real-time PCR and data analysis for the occludin gene expression. Data were analyzed based on clinical staging of breast cancer patients which were cited in data bank of TBI. Results: Results of this study have demonstrated that the occludin gene expression was increased with the advanced stage. In 22 of patients the expression of gene was elevated compared with normal samples. On the other hand, the expression was significantly increased in stage II in comparison with stage I. Conclusion: The expression of occludin has increased by elevation of stage compared with normal tissue. It is suggested that alteration in the expression of this gene might be a possible factor which could affect on patient’s prognosis the same as other factors which are belonging to the same family. Increasing in expression of this gene might be considered as one of the possible markers which predict the possibility of invasion and metastasis.

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Background: Staphylococcus aureus is the most important cause of nosocomial infections acquired in the community. Protein A is a major component of Staphylococcus aureus cell wall. In analysis of the nucleotide sequence Protein A encoding spa, locus x consists of 24 base pairs which repeat with high polymorphism. In this study, the spa gene of Staphylococcus aureus isolated from clinical specimens were obtained from patients admitted to the hospital and healthy carriers. Methods: In a cross-sectional study, a total of 200 samples were collected. One hundred fifty samples were obtained from hospitalized patients and 50 samples obtained from staff nasal swabs in Hamadan University Hospitals from October 2013 to August 2014. Disk diffusion antibiotic susceptibility tests performed. The antibiotics studied were Vancomycin (30 µg), Cefoxitin (15 µg) Gentamicin (10 µg), Tetracycline (30 µg), Trimethoprim/sulfamethoxazole (25 µg), Ciprofloxacin (5 µg), Erythromycin (15 µg), Clindamycin (2 µg), Rifampin (5 µg). The tests performed according to the guidelines of clinical and laboratory standards institute (CLSI). It also detect the mecA gene of Methicillin-resistant Staphylococcus aureus strains (MRSA) and genes spa which encodes the protein A by polymerase chain reaction (PCR). The PCR products using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method with enzyme Rsa I (Afa I) were prepared. Results: This methicillin-resistant Staphylococcus aureus strain (MRSA) had the highest sensitivity and resistance to ciprofloxacin and clindamycin. Totally, 8 amplicon with different sizes for the spa gene were identified. A total of 9 patterns polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) were found. Some of these patterns between Staphylococcus aureus isolated from clinical specimens and nasal carriers were common. Conclusion: There is a similar pattern of spa gene among patients admitted to the hospital and staff, according to our findings. Analysis of the patterns can reduced transmission of infection in both hospital staff and patients. Also it can help the physicians for correct management of infections.

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Background: Medullary thyroid cancer (MTC), includes 5-10% of all the thyroid cancers. RET proto-oncogene mutations have been found in association with MTC development. Therefore, identification of the mutations in RET can allow early diagnosis of the families who are at the risk of the disease. The goal of this study was to investigate existence and association between mutations in exon 19 of the RET proto-oncogene in an Iranian population medullary thyroid cancer patients and their family members. Methods: This study was run in the research laboratory of Research Institute for Endocrine Research Center Shahid Beheshti University of Medical Sciences from May, 2013 to May, 2014. In this study, 110 patients with confirmed medullary thyroid carcinoma were selected and examined. At first, the genomic DNA content of the peripheral white blood cells (WBC) of the samples were extracted using a saturated salting out and proteinase K standard method. Exon 19 of the RET proto-oncogene using polymerase chain reaction (PCR) method was amplified. Then the desired PCR products formation was confirmed by electrophoresis technique for true amplification, and finally the amplified samples were used for direct sequenced for finding and assessing any possible mutations Results: In this study, two nucleotide changes at position rs2075912 (Y: T/C) and position rs2075913 (W: T/A) exon 19 RET proto-oncogene were found in the patients with medullary thyroid cancer. The frequency of both nucleotide changes were higher in men than women with medullary thyroid cancer. The frequency of the rs2075912 and rs2075913 were 11.2 and 6.3% higher in men than women. But in statistical analysis, there was no association between age, sex and the founded two mutations. Conclusion: In addition to mutations in other exons of proto-RET, mutations in exon 19 can also be used for early detection and confirmation of medullary thyroid carcinomas.

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Background: Breast cancer is the most common non- skin cancer among women and it’s the second leading cause of cancer related death in women. Ubiquitin and ubiquitin like proteins are member of signal transduction pathways which have several cellular functions. It has shown that Ubiquitin like protein D (UBD) has accelerated the cancer progress. The aims of this study is evaluation of UBD gene expression in women suffering from breast cancer and its correlation with disease progression. Methods: In this study 30 FFPE (Formalin-fixed, paraffin-embedded) samples 20 cases from breast cancer and 10 cases from mammoplasty were collected from Parsian and Kasra Hospitals in Tehran after confirmation by pathologist. For each sample collection characters included ER-positive, lymph node negative, tumor size less than 5 cm in diameter were considered. Samples belonged to May 2010 up to April 2012. At first paraffin was removed by adding xylene then xylene removed with replacing ethanol 98%. After removing ethanol, RNA was extracted from samples by using RNX plus solution and cDNA synthesis were performed by using Moloney murine leukemia virus (M-MuLV) enzyme. UBD gene expression were examined in all samples cDNA by relative Real Time PCR. In this study GAPDH gene expression was also used as internal control. Results: UBD gene expression was obtained by calculating ΔΔCT and RQ. The average incensement of UBD gene expression in comparison of normal samples was 11 times. The results have shown that the level of UBD expression was related to the development and extend of the disease. In patients with stage 1 of disease, UBD gene expression had 2.73 times increase (P=0.001) compared to the control samples. However in stage 4 of disease, this number has increased up to 19.4 times (P=0.0005) more than normal. Conclusion: Considering the results of this study, it could be said that UBD gene expression as useful biomarker has an important role in detection of breast cancer. In addition as UBD gene expression levels increased stages of disease increased too. So that evaluation of UBD gene expression can be useful in early detection of disease.

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Background: Bloodstream infections (BSI) have a high incidence and high mortality in the worldwide. The mortality rate is variable between 20-70%. Therefore, early and timely detection of BSI agent in clinical laboratories is necessary. The aim of this study was to determine an efficient diagnostic tool to septicemia in accompany of blood culture method by Real-time PCR (using panbacterial 23S rRNA gene).

Methods: This cross-sectional study was conducted in two analytical and clinical stages in Hamadan University of Medical Sciences, Iran, from October 2014 to June 2015. In analytical stage, sensitivity (by serial dilution from 104 to 1 CFU/ml) and specificity of the primer were evaluated with the Staphylococcus aureus (as Gram positive indicator bacteria) and Escherichia coli (as Gram-negative indicator bacteria), human genome (from Hella cell culture), Candida albicans yeast and Aspergillus fumigatus fungus. In clinical stage, 121 blood samples were collected from patients suspected to sepsis in intensive care unit (ICU) from Hamadan University Hospitals. Finally, the results of Real-time PCR and blood culture methods were compared.

Results: The Real-time PCR showed a sensitivity ranging from 2 to 10 target copies per reaction to the whole blood for Escherichia coli and Staphylococcus aureus respectively. The specificity of this method was evaluated and no false positive amplification was identified. 57.85% (70 cases) of the samples were positive by Real-time PCR and 13.22% (16 cases) of the samples were positive by blood culture. However, none of the cases that were positive by blood culture were negative in Real-time PCR. As well as, 44.62% (54 cases) of cases were positive by Real-time PCR but blood culture showed no bacteria in the samples, and 42.15% (51 cases) were negative by both methods. Correlation or agreement of Kappa was 0.20, that indicating poor agreement between the two methods.

Conclusion: Real-time PCR is more sensitive than blood culture and also, because of high sensitivity of this primer by Real-time PCR, we can use it for screening blood samples from suspected patients of sepsis.

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Background: Over the last two decades invasive candidiasis has become an increasing problem in neonatal intensive care units (NICUs). Colonization of skin and mucous membranes with Candida spp. is important factor in the pathogenesis of neonatal infection and several colonized sites are major risk factors evoking higher frequencies of progression to invasive candidiasis. The aim of this study was to detect Candida colonization in NICU patients.

Methods: This cross-sectional study was conducted on 93 neonates in NICUs at Imam Khomeini and Children Medical Center Hospitals in Tehran. Cutaneous and mucous membrane samples obtained at first, third, and seventh days of patients’ stay in NICUs during nine months from August 2013 to May 2014. The samples were primarily cultured on CHROMagar Candida medium. The cultured media were incubated at 35°C for 48h and evaluated based on colony color produced on CHROMagar Candida. In addition, isolated colonies were cultured on Corn Meal Agar medium supplemented with tween 80 for identification of Candida spp. based on their morphology. Finally, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was performed for definite identification of isolated species.

Results: Colonization by Candida spp. was occurred in 20.43% of neonates. Fifteen and four patients colonized with one and two different Candida spp., respectively. Isolated Candida spp. identified as; C. parapsilosis (n: 10), C. albicans (n: 7), C. tropicalis (n: 3), C. guilliermondii (n: 2), and C. krusei (n: 1). In present study non-albicans Candia species were dominant (69.56%) and C. parapsilosis was the most frequent isolate (43.47%). Using Fisher's exact test, the correlation between fungal colonization with low birth weight, low gestational age, and duration of hospital stay was found to be statistically significant (P=0.003).

Conclusion: The results of this study imply to the candida species colonization of neonates. Neonates in NICU are at the highest risk for severe infection with Candida parapsilosis. Therefore, isolation of C. parapsilosis as the most common species (43.47%) in present study was noteworthy.