Development of a multiplex real-time PCR assay for the detection of Treponema pallidum, HCV, HIV-1 and HBV.

This study aims to establish an assay to detect these pathogens in clinical serum specimens. We established a TaqMan-LNA real-time PCR assay for rapid, sensitive, specific, quantitative, and simultaneous detection and identification. The copy numbers of standards of these four pathogens were quantified. Standard curves were generated by determining the mean cycle threshold (Ct) values versus 10-fold serial dilutions of standards over a range of 10(6) to 10(1) copies / μL with the lowest detection limit of the assay at 10(1) copies / μL. The assay was applied for 328 clinical specimens, and compared with ELISA and commercial NAT method. The assay identified 39 T. pallidum-, 96 HCV-, 13 HIV-1-, 123 HBV-, 5 HBV/HCV-, 1 T. pallidum/HBV-,1 HIV-1/HCV-, 1 HIV-1/T. pallidum- positive specimens. By showing greater sensitivity, this assay becomes a highly reliable, cost-effective and useful molecular diagnostic tool for large-scale screening of clinical specimens and will assist in the study of the pathogenesis and epidemiology of sexually transmitted blood diseases. PMID: 25866106 [PubMed - as supplied by publisher]
Source: Japanese Journal of Infectious Diseases - Category: Infectious Diseases Authors: Tags: Jpn J Infect Dis Source Type: research