荧光定量PCR法在丙型肝炎病毒核糖核酸检测中的应用

Application of fluorescent quantitative PCR in detection of HCV RNA

目的：通过与荧光半定量PCR及传统逆转录PCR（RT-PCR）比较,探讨荧光定量PCR（FQ-PCR）在丙型肝炎病毒核糖核酸（HCV RNA）检测中的应用价值。方法：采用FQ-PCR、荧光半定量PCR及RT-PCR三种方法同步检测180例丙型肝炎患者HCV RNA含量,所得结果采用χ^2检验。结果：FQ-PCR在血浆中及单个核细胞中的阳性检出例数最多,分别占78.3%和83.7%。无论是在血浆中或单个核细胞测定,FQ-PCR阳性检出率均明显高于同步检测的荧光半定量PCR及RT-PCR（P〈0.01）。以RT-PCR为标准,当HCV RNA水平在10^3～5copies/ml时,FQ-PCR和荧光半定量PCR阳性检出水平相一致（P〉0.05）,明显高于RT-PCR（P〈0.01）。当HCV RNA水平在10^6～7cop-ies/ml时,3种检测方法的阳性检出率基本相同（P〉0.05）。结论：FQ-PCR检测HCV RNA具有高灵敏度、高特异性,且二次污染的可能性小。

Objective： To discuss the application value of fluorescent quantitative PCR （FQ-PCR） in detection of HCV RNA by comparing fluorescent seniquantitative PCR and conventional real time PCR （RT-PCR） . Methods： FQ-PCR, fluorescent semiquantitative PCR and RT-PCR were used simultaneously to detect the copy level of HCV RNA in 180 HCV patients. The data were calculated by χ^2 test. Results： FQ-PCR could detect the largest number of positive cases in plasma and mononuclear cells accounting for 78.3% and 83.7% respectively. The positive rate of FQ-PCR method was significantly higher than fluorescent semiquantitative PCR and RT-PCR method （P 〈0. 01 ） in either plasma or mononuclear cells. When the copy numbers of HCV RNA was between 10^3-5/ml, FQ-PCR and semiquantitative PCR methods had similar positive detection rate （ P 〉 0.05 ）, but they had higher sensitivity than RT-PCR （P 〈0. 01 ） . The above three detection methods had similar positive detection rates （P 〉 0. 05 ） when the copy numbers of HCV RNA was 10^6-7/ml. Conclusion： FQ-PCR method has high sensitivity and specificity in detection of HCV RNA, and the likelihood of reagent pollution in FQ-PCR method is small.