Multiplex fluorescence quantitative polymerase chain reaction for simultaneous detection of hepatitis B virus, hepatitis C virus and human immunodeficiency virus.

Background: To construct a multiplex fluorescence quantitative polymerase chain reaction (MFQ-PCR) system to simultaneously detect hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) cDNA, and human immunodeficiency virus (HIV) cDNA to reduce the false positive and false negative results, therefore increasing the efficacy and accuracy of donor blood testing. The feasibility of the MFQ-PCR system was assessed by testing 58 clinical serum samples from hospitalized patients.

Methods: TaqMan probes designed specifically for the detection of HBV, HCV, and HIV viruses separately were put into the same MFQ-PCR tube and co-amplified with the HBV target DNA, HCV target cDNA, and HIV target cDNA.

Results: In this new MFQ-PCR system, the detection limits were about 10 copies for each of standard plasmid DNAs of HBV, HCV, or HIV. In the test of 58 clinical serum samples, of which 23, 16, and 3 were positive for HBV, HCV, and HIV, respectively, by ELISA method, and 16 were normal controls, this new MFQ-PCR system detected 41 of 42 positive samples, and the 16 normal controls were all negative. The rate of consistency between the new system and ELISA was 98.3%.

Conclusions: MFQ-PCR has the advantage of simultaneous detection of HBV, HCV and HIV viruses in clinical serum samples with high efficiency and accuracy.