荧光定量PCR技术在检测HCV—RNA中的应用

目的 评价荧光定量ＰＣＲ技术测定ＨＣＶ－ＲＮＡ水平的价值，探讨ＰＢＭＣ中检出ＨＣＶ－ＲＮＡ的意义。方法 采用荧光定量ＫＲ及ＲＴ－ＰＣＲ技术同时检测８７例血液透析患者的血清和淋巴细胞中ＨＣＶ－ＲＮＡ。结果血清、淋巴细胞中ＨＣＶ－ＲＮＡ的荧光定量ＰＣＲ检出率（４９．４％、６９．０％），分别高于相应标本中ＨＣＶ－ＲＮＡ的ＲＴ－ＰＣＲ阳性率（３３．３％、３５．６％）（Ｐ ＜０．０５）。血清、淋巴细胞中同时检出ＨＣＶ－ＲＮＡ的一致率，荧光定量ＰＣＲ法（３１．０％）显著高于ＲＴ－ＰＣＲ定性法（６．９％）（Ｐ＜０．００１）． 荧光定量ＰＣＲ法，淋巴细胞中ＨＣＶ－ＲＮＡ检出率高于血清（Ｐ＜０．０１）。结论 与 ＲＴ－ＰＣＲ相比，荧光定量ＰＣＲ技术检测 ＨＣＶ－ＲＮＡ敏感性较高，对ＰＢＭＣ内ＨＣＶ－ＲＮＡ检测有利于提高ＨＣＶ检测的阳性率。

Application of amplisensor quantitative PCR in detecting HCV-RNA

Objective The aim of the paper is to study the meaning of HCV-RNA in peripheral blood mononuclear cell (PBMC)and to evaluate the technique of amplisensor quantitative PCR in detecting HCV-RNA. Methods Serum and PBMC of 87 hemodialysis patients were tested for hepatitis C viral RNA by amplisensor quantiative PCR and RT-PCR (qualitative) Results Positive rate of HCV - RNA in serum and I'BMC by amplisensor quantitative PCR(49. 4% 69.0% )was significantly higher than by RT-PCR respectively(33. 3% .35. 6% ) ( P <0. 05). The rate of simultaneous positive HCV-RNA in serum and PBMC was significartiy higher by amplisensor quantitative PCR (31. 0%) than by RT-PCR (6. 90% ) ( P <0.001). Positive rate of HCV-RNA in PBMC was significantly higher than in serum by amplisensor quantiative PCR ( P <0.01). Conclusion Compare with RT-PCR, amplisensor quantitative PCR technique has higher sensitivity in delecting HCV-RNA. Detection of HCV-RNA in PBMC will contribute to improve the diagnosis of hepatitis C viral.