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Title

Evaluation of the Performance Characteristics of an In-House One Step TaqMan Real Time-Polymerase Chain Reaction Assay for Detection and Quantification of Hepatitis C Virus

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Abstract

 Background: With theimprovementof quantitative molecular hepatitisCvirus (HCV)RNAassays, the usefulness of these assays has been indicated for management of HCV infection. Recently, various real time assays with different methodology and performance characteristics have been introduced. Objectives: This study aimed at designing, developing, and evaluating an in-house 1 step TaqMan real time-polymerase chain reaction (RT-PCR) assay for detection and quantification of HCV-RNA. Methods: The primers and Probe were selected from a highly conserved region of the HCV genome, which allowed the detection of 4 common HCV genotypes in Iran. Using 4 quantification standards from 101 IU/ L to 104 IU/ L and clinical specimens, the current study determined analytical sensitivity, linear range, precision, analytical and clinical specificity, and validity of the assay. Data was analyzed by the SPSS statistical software (version 16). Results: The sensitivity of the assay with 95% probability, determined by probit analysis, was 15 IU/ L. The assay showed a linear range of 101 IU/ L to 104 IU/ L (R2 = 0. 989). The coefficient of variation for intra and inter assay precision of the assay, based on threshold cycle value, ranged from 0. 24 to 0. 4, and from 1. 94 to 3. 19, respectively. The analytical and clinical specificity was 100%. No bias in relation to concentration between the results of 29 HCV RNA positive clinical specimens, simultaneously tested by artus HCV LC RT-PCR reagents and in-house reagents, was observed in the method comparison. Conclusions: The in-house 1 step TaqMan Real Time RT-PCR assay showed acceptable performance characteristics. Our study presents the robustness and cost-effectiveness of the method for detection and quantification of HCV RNA.

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    APA: Copy

    Ranjbar Kermani, Fahimeh, Amini Kafi Abad, Sedigheh, MOUSAVI HOSSEINI, KAMRAN, Maghsoudlu, Mahtab, SHARIFI, ZOHREH, & SAMIEE, SHAHRAM. (2017). Evaluation of the Performance Characteristics of an In-House One Step TaqMan Real Time-Polymerase Chain Reaction Assay for Detection and Quantification of Hepatitis C Virus. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), 10(3), 0-0. SID. https://sid.ir/paper/317690/en

    Vancouver: Copy

    Ranjbar Kermani Fahimeh, Amini Kafi Abad Sedigheh, MOUSAVI HOSSEINI KAMRAN, Maghsoudlu Mahtab, SHARIFI ZOHREH, SAMIEE SHAHRAM. Evaluation of the Performance Characteristics of an In-House One Step TaqMan Real Time-Polymerase Chain Reaction Assay for Detection and Quantification of Hepatitis C Virus. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM)[Internet]. 2017;10(3):0-0. Available from: https://sid.ir/paper/317690/en

    IEEE: Copy

    Fahimeh Ranjbar Kermani, Sedigheh Amini Kafi Abad, KAMRAN MOUSAVI HOSSEINI, Mahtab Maghsoudlu, ZOHREH SHARIFI, and SHAHRAM SAMIEE, “Evaluation of the Performance Characteristics of an In-House One Step TaqMan Real Time-Polymerase Chain Reaction Assay for Detection and Quantification of Hepatitis C Virus,” JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), vol. 10, no. 3, pp. 0–0, 2017, [Online]. Available: https://sid.ir/paper/317690/en

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