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Báo cáo y học: " A novel duplex real-time reverse transcriptase-polymerase chain reaction assay for the detection of hepatitis C viral RNA with armored RNA as internal control"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: A novel duplex real-time reverse transcriptase-polymerase chain reaction assay for the detection of hepatitis C viral RNA with armored RNA as internal control | Meng and Li Virology Journal 2010 7 117 http www.virologyj.eom content 7 1 117 VIROLOGY JOURNAL RESEARCH Open Access A nlovel duplex real-time reverse transcriptase-polymerase chain reaction assay for the detection of hepatitis C viral RNA with armored RNA as internal control Shuang Meng 1 2 and Jinming Li 1 2 Abstract Background The hepatitis C virus HCV genome is extremely heterogeneous. Several HCV infections can not be detected using currently available commercial assays probably because of mismatches between the template and primers probes. By aligning the HCV sequences we developed a duplex real-time reverse transcriptase-polymerase chain reaction RT-PCR assay using 2 sets of primers probes and a specific armored RNA as internal control. The 2 detection probes were labelled with the same fluorophore namely 6-carboxyfluorescein FAM at the 5 end these probes could mutually combine improving the power of the test. Results The limit of detection of the duplex primer probe assay was 38.99 lU ml. The sensitivity of the assay improved significantly while the specificity was not affected. All HCV genotypes in the HCV RNA Genotype Panel for Nucleic Acid Amplification Techniques could be detected. In the testing of 109 serum samples the performance of the duplex realtime RT-PCR assay was identical to that of the COBAS AmpliPrep CAP COBAS TaqMan CTM assay and superior to 2 commercial HCV assay kits. Conclusions The duplex real-time RT-PCR assay is an efficient and effective viral assay. It is comparable with the CAP CTM assay with regard to the power of the test and is appropriate for blood-donor screening and laboratory diagnosis of HCV infection. Background Hepatitis C virus HCV is one of the major causes of chronic liver diseases which has infected an estimated 170 million people worldwide 1 2 . It is responsible for chronic liver diseases and is a risk factor for liver cirrhosis and hepatocellular carcinoma 3 . Early diagnosis and evaluation of HCV cases is very .