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Showing 2 results for Real-Time Pcr
Samira Sheikh Ghomi , Parisa Farnia , Mojtaba Darbouy ,
Volume 14, Issue 2 (7-2014)
Abstract
Background & objectives: The rapid identification of patients carrying resistant Mycobacterium tuberculosis (M.TB) isolates is important for effective tuberculosis therapy. Unfortunately, during the recent years considerable numbers of isolates showed resistant to Rifampin (RIF) and Isoniazid (INH). The aim of this study was to rapidly identify resistant MTB isolates using molecular method. For this reason, the comparison between real-time PCR based on Taqman and HRM AssayS in detection of rpoB, inhA and katG genes mutation in clinical isolates were performed and analyzed. Methods: The study carried out on Mycobacteriology Research Center (MRC) from 2012-2013. Classical susceptibility testing i.e., proportional method against INH and RIF was performed on eighty three M.TB isolates. Thereafter, multiplex and real-time PCR were performed on extracted DNA sample. The real-time PCR was based on Taqman and HRM assays. Mutation in genes rpoB, inhA and katG were detected. Results: In overall, based on proportional and multiplex PCR method, 47 and 35 isolates were resistant to RIF and INH, respectively. Thirty of strains were resistant to both RIF and INH. The agreement of real-time PCR using Taqman was 88% for resistant and 84% for susceptible isolates, whereas the agreement of HRM was 96% and 30%, respectively. The sensitivity and specificity of Taqman in comparison to multiplex were 84% and 88%, respectively. In addition, the sensitivity and specificity of HRM were 30% and 96%, respectively. Conclusion: Results documented that real-time PCR based on Taqman assay is more sensitive than HRM assay. Additionally, real-time PCR based on Taqman assay is a rapid, accurate and cost effective method in detection of Mycobacterium tuberculosis resistance.
Khadijeh Khanaliha, Farah Bokharaei-Salim, Mohsen Sadeghi, Borna Salemi,
Volume 22, Issue 3 (10-2022)
Abstract
Background & objectives: Toxoplasma gondii is an obligate intracellular parasite with global distribution. Diagnosis of Toxoplasma gondii infection with high sensitivity and specificity is very important in managing and treating this disease. The purpose of this study is serological and molecular investigation of toxoplasmosis using B1 gene in HIV- positive patients referred to hospitals affiliated with Iran University of Medical Sciences.
Methods: In this study, 660 blood samples were collected from HIV/AIDS- positive patients referred to hospitals affiliated with Iran University of Medical Sciences. Patient samples were examined for the presence of IgG and IgM antibodies against Toxoplasma gondii using an ELISA kit. Genomic DNA was extracted from the patient's serum, as well as peripheral blood mononuclear cell (PBMC) and whole blood samples, and then Real time-PCR was performed.
Results: Although IgG antibody against Toxoplasma gondii was positive in 158 (23.9%) patients out of 660 HIV- positive patients, IgM antibody was positive in 5 (0.76%) patients. The results of Real-Time PCR showed that 7 (1.06%) patients were positive in PBMC samples, of which five patients were positive for IgM antibodies against Toxoplasma gondii while two patients had high- level Toxoplasma IgG antibody titers.
Conclusion: The results of the study indicate that the Real-time PCR method using PBMC DNA samples is a suitable method for the diagnosis of toxoplasmosis. This method, together with the antibody test, especially the high titer of Toxoplasma IgG antibodies, can be helpful in the diagnosis of toxoplasmosis.
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