PCR-BHEL技术的建立及对血清中丙型肝炎病毒的检测
摘要
目的聚合酶链反应偶联印迹杂交和酶联放大技术(PCR-BHEL)的建立并应用该技术检测血液传染性病毒。方法综合聚合酶链反应的高敏感性及印迹杂交和酶联免疫技术的高特异性建立起PCR-BHEL技术,然后应用该技术对酶联免疫吸附实验(ELISA)检测的174例血清标本(其中129例抗-HCV阳性,45例抗-HCV阴性)的HCV RNA进行检测,并将该技术与逆转录聚合酶链反应(RT-PCR)及荧光定量聚合酶链反应(FQ-PCR)HCV RNA的检测结果进行比较,以确定该技术在检测血液传染性病毒中的优越性(高特异性、高敏感性)。结果129例抗-HCV阳性标本经PCR-BHEL和RT-PCR检测HCV RNA的检出率分别为78.29%(101/129)和62.02%(80/129);45例抗-HCV阴性标本经PCR-BHEL和RT-PCR检测HCV RNA的检出率分别为6.67%(3/45)和2.22%(1/45),两种检测方法的差异有统计学意义(0.010.05)。结论PCR-BHEL较RT-PCR的灵敏度高,能够有效地提高HCV RNA的检出率,可推广应用于其他血液传染性病毒的检测。
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(0.01m samples were 75.82%(69/91)and 62.64%(57/91)when they were detected by PCR-BHEL and FQ-PCR respectively, the HCV RNA results of 18 anti-HCV negative serum samples were all negative when they were detected by the two methods, the difference has no statistic differences(P>0.05). 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.
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(0.01
引文
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