Establishment Of PCR-BHEL Technology And Detection Of Hepatitis C Virus In Serum

Objective To establish polymerase-chain reaction coupled with blot-hybridization and enzyme-linked technology (PCR-BHEL) for HCV detection in serum and use this technology to detect infectious virus in blood. Methods The PCR-BHEL technology which was based on the high sensitivity of PCR and the high specificity of blot-hybridization and enzyme immunoassay has been established. Then using this technology detected HCV RNA of 129 anti-HCV positive and 45 anti-HCV negative serum samples (detected by ELISA). After 174 serum samples were detected by PCR-BHEL and reverse transcription polymerase chain reaction (RT-PCR)/fluorescence quantitative polymerase chain reaction (FQ-PCR) respectively, compared the HCV RNA results of PCR-BHEL with RT-PCR /FQ-PCR respectively to confirm the superiority (high sensitivity and high specificity) of PCR-BHEL in the detection of infectious virus in blood. Results The HCV RNA positive rates of 129 anti-HCV positive serum samples were 78.29%(101/129) and 62.02%(80/129) when they were detected by PCR-BHEL and RT-PCR respectively, the HCV RNA positive rates of 45 anti-HCV negative serum samples were 6.67%(3/45) and 2.22%(1/45) when they were detected by PCR-BHEL and RT-PCR respectively, the difference has statistic significance(0.010.05). Conclusion PCR-BHEL is obviously superior to RT-PCR, and higher than RT-PCR in sensitivity. The results imply that PCR-BHEL is a sensitive method and can be applied in the detection of other infectious viruses in blood.