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Background and Objective: Sexually infections transmitted by bacteria are one of thetherapeutic and social problemsworldwide. The Real-time PCR assay is one of the most sensitive diagnostic and screening methods for these infections. The purpose of this study wassimultaneous detection of Chlamydia trachomatis and Mycoplasma genitaliumusing the TaqMan duplex real-time polymerase chain reaction. Materials and Methods: In the present study, we used extracted DNA from women with vaginal infections available in the bank ofthe molecular and medicalresearch center, Arak University of Medical Sciences, Arak, Iran. The duplex real-time PCR was evaluated to detectChlamydia trachomatis and Mycoplasmagenitalium in 110 vaginal andendocervical samples. To validate the diagnostic performance of the designedmethod, the tests were compared with a commercial diagnostic kit. Results: The size of the ampliconsOmpA, 135 bp and MgPa, 145bp,indicated the accuracy of the primers designing procedures. The analysis of a panel of human STI revealed no cross-reactivity in the method. The diagnostic sensitivity and specificity of developedTaqMan duplex real-time PCR were 97% and 75%, respectively,compared to a commercial diagnostic kit. Conclusion: The results indicated that the used TaqMan duplex real-time PCR technique is specific and cost-effective and can be an excellent alternative to commercial kits for simultaneously detecting C. trachomatis and M. genitalium. Although the disadvantage of this method is its high cost, this disadvantage has significantly been resolved by the multiplex method.

Background and Aim: Cervical Cancer (CC) is the third most common malignancy in the women, the main cause of which is human papillomavirus (HPV). Both E6 and E7 oncogenes of the virus play an important role in its tumorigenesis. Today, methods available for screening CC are not capable of detecting the disease at an early stage. Therefore, it is important to identify new biomarkers for early detection of this cancer. For this purpose, in the present study, miRNAs targeting the two oncogenes E6 and E7 of human papillomavirus (types 16 and 18) were studied in CC by bioinformatics. Methods & Materials: First, using the NCBI database, the E6 and E7 gene sequences were obtained for both human papillomavirus types 16 and 18. Then, using the miRBase and RNA22 bioinformatics databases, the most appropriate targeting miRNAs for these genes were selected. Ethical Considerations: This study was approved by Ethics Committee of Arak University of Medical Sciences. Results: Based on the P obtained from bioinformatics databases, miRNA including miR-92a-5p (P=7.51e-2), miR-195-3p (P=2.24e-1), miR-34a-5p (P=2.73e-1) and miR-155-5p (P=4.95e-2) were introduced for the two genes E6 and E7. Conclusion: Results from bioinformatics studies revealed that of the four miRNAs identified, miR-155-5p and miR-92a-5p are probably the targeting miRNAs specific for the E6 and E7 genes, respectively. Therefore, it seems that these miRNAs can be a suitable candidate for in vitro studies in CC patients.

Background and Aim Chronic Lymphocytic Leukemia (CLL) is the most commonly occurring leukemia in adults, accounting for about 30-25% of total leukemia. One of the important etiological causes of this leukemia is the disruption of the Nuclear Factor Kappa B (NF-kB) signaling pathway. The two proteins of Apoptosis-Inducing Ligand (APRIL) and B-Cell Activating Factor (BAFF) play a role in the pathogenesis of this leukemia by affecting the NF-kB signaling pathway. In this study, due to the effect of miRNAs in regulating many cellular processes, the prediction of the prominent miRNAs targeting APRIL and BAFF transcripts in B-cell CLL patients was evaluated using specific and different bioinformatics programs. Methods & Materials Afterwards retrieving the sequences of APRIL and BAFF proteins from the NCBI website, by using several programs including miRanda, TargetScan, miRWalk, DIANA and miRDB with different algorithms, the prediction of miRNAs targeting these genes was investigated. Ethical Considerations This study was approved by the Research Ethics Committee of Arak University of Medical Sciences. Results Based on the scoring system of bioinformatics programs, “hsa-miR-145-5p” and “hsa-miR-185-5p” were identified as miRNAs targeting APRIL gene, while “hsa-miR-424” and “hsa-miR-497”were miRNAs targeting BAFF gene. They were suggested for the practical studies in future. Conclusion Based on the important role of APRIL and BAFF genes in the normal process of cell death and B-cell evolution, it seems that the mi-RNAs predicted by bioinformatics programs using different algorithms can be used as a diagnostic molecular biomarker to identify B-cell CLL patients.

Background: Campylobacter genus is considered some of the most important agents of bacterial gastroenteritis worldwide. Campylobacter coli (C. coli) is accounted to at least 25% of all Campylobacter related diarrheal diseases moreover, C. coli infections can result in severe complications, such as bacteremia, sepsis, meningitis and spontaneous abortion. Finally, there is evidence that the frequency of antimicrobial resistance is higher in C. coli, when compared to C. jejuni. There is no data regarding the frequency and antibiotic resistance profile of C. jejuni isolated from human gastroenteritis samples. The present study aimed to determine the frequency and antibiotic resistance patterns of Campylobacter coli isolated from infectious diarrhea samples. Methods: In a descriptive cross-sectional study, 200 infectious diarrhea samples collected in Arak University of Medical Sciences Hospitals, Markazi Province, Iran, from May to November 2015 were subjected to the study. In order to identify C. coli modified Gram stain, modified charcoal-cefoperazone-deoxycholate agar (mCCDA) and Brucella agar media with filter and CeuE gene polymerase chain reaction (PCR) were accomplished. Antibiotic resistance against tetracycline, erythromycin, ciprofloxacin, ampicillin and gentamicin was evaluated phenotypically and genotypically. Results: In total, out of 200 modified gram stained samples, 2 cases (1%) of C. coli were identified. Cultivating methods using mCCDA medium found 2 isolates (1%), 3 isolates (1.5%) were grown on Brucella agar with filter and 5 cases (2.5%) were determined as C. coli using PCR assay. Antibiotic resistance was observed in 5 cases against tetracycline, erythromycin and ciprofloxacin (100%), in 4 cases against ampicillin (80%), in 2 cases against gentamicin (40%), in 5 cases with CmeB, 23srRNA mutation in, qnrS, tet (o) (100%), in 4 cases with gyrA4 (80%), in 3 cases with gyrA5 (60%), in 5 cases with gyrA6 (100%), in 4 cases with Oxa61 (60%) and in 1 case with aphA-3-1 (20%). Conclusion: In this present study C. coli with low prevalence and entire resistance to ciprofloxacin and erythromycin which are the first line antibiotic for the treatment of campylobacter gastroenteritis is introduced as a causative agent of gastroenteritis in patients at central part of Iran.

Abstract Background: Hepatocellular carcinoma (HCC) is the third major cause of cancer death worldwide. Hepatitis B virus (HBV) and HBx gene play an important role in the development of HCC by influencing signaling pathways. Since there is no detectable symptom in the early phase of HCC, there is need to find new HCC-specific markers with high sensitivity for early detection and diagnosis of HCC. On the other hand, by the advent and development of bioinformatic sciences, it is now possible to predict miRNAs as biomarkers, and their targets. Therefore, in the present study, based on the results of the bioinformatic software applications with different algorithm, we selected the miRNA targeting HBx and NOTCH1 mRNAs according to higher score, suitable connection with target gene and confirming them in more softwares. Materials and Methods: First, the sequences of NOTCH1 and HBx genes were retrieved from NCBI. Afterwards, several software applications such as TargetScan, mirWalk, miRBase, Miranda, PicTar, miRVir, and DIANA were applied to predict miRNAs. Results: Based on the high scoring by bioinformatics softwares and suitable targeting, miR-34a were selected to target NOTCH1 and miR-6510, miR-5193 and miR-214 were chosen to targetHBX gene. Conclusion: Because of tumor suppression roles of miR-214 and miR-34a, they probably could be used as therapeutic strategy in cancer researches. It is also seems that the miR-5193 could act as a specific marker in Hepatocellular carcinoma.

Abstract Background: Ureaplasma and M. genitalium species belong to a kind of bacteria that are sexually transmitted and are the possible cause of pelvic inflammatory disease and nongonococcal urethritis, and et al. The aim of this study was to determine the urea plasma and Mycoplasma genitalium species frequency in women with vaginal infection and various sexual partners who referred to women, s health promotion and treatment center in Arak. Materials and Methods: Endocervical swab samples from 110 women with vaginal infections referred to women’s health promotion and treatment center in Arak, were prepared. Patients’ personal information and identities during reception process were registered. The samples were transferred to the laboratory in the transport environment and after DNA extraction, were evaluated according to Real-time PCR assay. Results: Urea plasma and Mycoplasma genitalium bacteria existed in 96(87.27%) and 4(3.63%) of patients, respectively. Among them, 4 cases had both bacteria infections. The amount of isolation in young women between 30-39 years old was more than others. Conclusion: The results show that the colonization of urea plasma species in adult women is 40-80% and in studied group is 87.27%. These results indicate that with due attention to the increasing number of sexual partners and the increase of sexual activity, the urea plasma colonization of women will increase. In view of the potential influence of mycoplasma species on side effects resulted from pregnancy infection of mothers and mortality, on-time diagnosis and treatment will be increasingly essential.

Abstract Background: Circulating microRNAs are promising biomarkers in diagnosis and assessment of cancerous patients. Quantitative Real-time PCR assay is a sensitive test for evaluating the levels of miRNAs expression. Nevertheless, there is no concurrence on selecting appropriate reference genes for qPCR analysis of miRNAs in circulation. Therefore, the current study aimed to select a suitable reference gene for normalizing the RT-qPCR assay results in plasma samples of patients with gastric cancer. Materials and Methods: Based on previously published studies, three molecules SNORD47, U6 RNA, and miR-103 were selected as the candidate reference genes. After RNA extraction from plasma samples of 40 patients with gastric cancer and 40 healthy individuals, expression levels of these molecules were evaluated using Real-time PCR method. Results: The results showed that the developed assays are able to diagnose their specified targets by a suitable linear range. By comparing patients and control groups, although the expression levels of miR-103 molecule were not equal between the two groups (p= 0.017), SNORD47 and U6 RNAs had similar expression levels. However, the variations of SNORD47 expression were lower that U6 RNA. Conclusion: Based on the results of the current study, the SNORD47 molecule has a stable expression levels in plasma samples of patients with gastric cancer and normal individuals and can be used as an appropriate reference gene for normalizing the quantitative data of qPCR assay.

Background: Due to some problems of serological assays in diagnosis of patients infected with HIV-1 and HCV, the use of molecular assay has become widely accepted. However, molecular assays have a major drawback of high cost and dependency to expensive thermocycler instruments. Methods: This research described the development of a multiplex nucleic acid sequence-based amplification (NASBA) assay for the simultaneous or single detection of HIV-1 and HCV genomes in plasma samples. The sensitivity and specificity of this assay has been evaluated using several clinical samples. Findings: The results showed that the primers used in this assay did not have any interactions with each other or with other possible interfering agents. The analytical sensitivity of this assay for both HIV-1 and HCV was calculated as 1000 copies/ml. The clinical sensitivity and specificity of the assay were considered as 90% and 100%, respectively. Conclusion: The developed multiplex NASBA assay can be used for simultaneous or single detection of HIV-1 and HCV viruses with a suitable sensitivity and specificity. Due to easy application, multiplexity, and no need to thermocycler instruments, this assay can be used as a low cost method in laboratories with limited instruments.