Background and Objectives: PCR is a sensitive assay and could be used as an accurate diagnostic method for detecting various types of microorganisms’ genome in low concentration in biological specimens. The demand for sensitive, rapid, safe and easy detection of PCR products has led researchers to a combination of this method with ELISA. Materials and Methods: Conserved sequences were selected for design of primers. Samples were tested by ELISA (for detection of specific HIV and HCV proteins) and real- time PCR for detection of specific nucleic acid and viral genome respectively. Viral genome was extracted and reverse transcription was performed with M-Mulv and the cDNA kept at -80°C. The PCR products were labeled by DIG-dUTP. Diluted PCR products were analyzed with both electrophoresis and ELISA methods.Results: Thirty-five samples were tested with the PCR-ELISA method. False positive or negative reactions were not observed. ELISA results of diluted products were compared with results obtained by electrophoresis. In gel electrophoresis, dilution of 1/10 was positive, but in ELISA, optical density of 1/100 dilution was much more than the cut-off value. Conclusion: Detection limits for gel electrophoresis as well as ELISA have been evaluated. It was shown that the PCR-ELISA method is ten times more sensitive than conventional PCR.

Introduction: Annually, more than 14 million people are reported to be infected with Leishmaniasis all over the world. In Iran, this disease is seen in the form of cutaneous and visceral leishmaniasis, of which the cutaneous form is more wide spread. In recent years, cutaneous leishmaniaisis is diagnosed by PCR utilizing specific primers in order to amplify different parasite genes including ribosomal RNA genes, kinetoplast DNA or tandem repeating sequences. The aim of this research was to detect early stage cutaneous leishmaniasis using Multiplex-PCR technique. Methods: In this study, 67 samples were prepared from patients with cutaneous leishmaniasis. DNA was extracted with phenolchloroform. Each specimen was analyzed using two different pairs of PCR primers. The sensitivity of each PCR was optimized on pure Leishmania DNA prior to use for diagnosis. Two standard parasites L. major and L. tropica were used as positive control. Results: DNA amplification fragments were two 115 bp and 683 bp for AB and UL primers, respectively. The sensitivity of two primers was not equal for detection of L. major and L. tropica. The sensivity of PCR with AB primer was 35 cells, while that for UL primer was 40 cells. Conclusion: The results of this study indicate that PCR is a sensitive diagnostic assay for cutaneous leishmaniasis and could be employed as the new standard for routine diagnosis when species identification is not required. However, the ability to identify species is especially important in prognosis of the disease and in deciding appropriate therapy, especially in regions where more than one type of species and disease are seen by clinicians.

BACKGROUND AND OBJECTIVE: Staphylococcus aureus is one of the most common causes of food poisoning in the world. Various studies indicated that 18-15 percent aureus strains isolated from different sources are be able to produce enterotoxin which known as the main factor of poisoning. The aim of this study was to identify enterotoxin genes, such as: (femA, see, sed, sec, seg, seb, sea) in S. aureus confirmed using multiple PCR method.METHODS: In this study, 60 samples of Staphylococcus aureus isolated from purulent infections, skin and have symptoms of poisoning: vomiting and diarrhea in humans, production of enterotoxin was examined. After DNA extraction of isolates, multiple PCR using specific primers for enterotoxin genes was performed.FINDINGS: Overall, 50% of Staphylococcus aureus isolates contain one or more enterotoxin gene. The most abundant gene was sea (30%) and sed (10%), see (3.8%), sec (6.1%) were also identified.CONCLUSION: It was found that other genes such as TSST-1 are involved staphylococcus aureus enterotoxin production in the creation of visual acuity in addition to above genes. In general, the presence of Staphylococcus aureus in human clinical specimens, especially enterotoxigenic strains, can be considered a potential risk for health

Mycoplasmas hominis, Mycoplasmas genitalium and Ureaplasma urealyticum are associated with infections of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. A Multiplex PCR was developed for simultaneously detection of theseMycoplasmas species in a single amplification reaction. The total number of 104 samples was collected from 104 women’s genital specimens with urogenital infections for identification of M.hominis, M.genitalium and U.realyticum by Multiplex PCR. The High Pure PCR Template Preparation Kit purified nucleic acids from 100 ml of specimen (American, Roche Company).In addition to the kit, boiling method was used to extraction of DNA from samples.UUA2 and UUS2 primers were used for urease gene amplification of U.urealyticum, MH1 and MH2 used for 16S rRNA gene amplification of M.hominis, and Adhesion protein gene (MgPa) used for 16S rRNA gene amplification of M.genitalium in all samples. The number of 28 samples (27%) was positive for Mycoplasmas. M.hominis, M.genitalium, and U.urealyticum were detected in 8.7, 3.9, and 14.5 percent of samples, respectively. The accumulated frequencies for M.hominis, M.genitalium, and U.urealyticum were 9 (8.7%), 4 (3.9%), and 15 (14.5 %), respectively. The results of this study revealed that Multiplex PCR is a highly sensitive, specific and cost-effective test for screening of genitourinary tract infections.