不同混合方式检测丙型肝炎病毒核酸的研究

刘沛; 王灿楠; 朱凤才; 张言超; 史志旭; 寇海兵

不同混合方式检测丙型肝炎病毒核酸的研究

Study on different mnipool methods for detection of hepatitis C virus nucleic acid

摘要

摘要:
目的研究不同混合方式检测丙型肝炎病毒核酸含量(HCV cDNA)的方法。
方法以未混合标本为对照, 使用荧光定量PCR(FQ-PCR)技术研究提取阶段混合, 反转录阶段混合以及提取和反转录阶段均混合等不同实验条件下的HCV cDNA含量。
结果提取阶段混合组的HCV cDNA含量与对照组相比未见显著性差异(P>0.05);反转录阶段混合组以及提取和反转录阶段均混合组的HCV cDNA含量低于对照组(P < 0.05), 且在低浓度时出现假阴性结果.阴性标本在提取阶段混合未见假阳性。
结论在对混合血清进行HCV cDNA测定时, 应将混合安排在提取阶段, 不宜为增大混合标本数而在提取和反转录阶段均混合。

Abstract:
Objective To develop the minipool technology for detection of hepatitis C virus nucleic acid as a screening strategy of molecular epidemiology.
Methods The use of fluorescence polymerase chain reaction determined the contents of hepatitis C virus cDNA in different minipool groups that minipools were formed in the stage of viral RNA extraction.The tests of individual sera were taken as the control group.
Results It was found that the content of HCV cDNA for extraction minipool group had no difference from that of the control group(P>0.05), but the content of HCV cDNA for reverse transcription group and extraction and reverse transcription group was lower than that of the control group(P < 0.05)and weak positive sample missed the detection.
Conclusion The minipool in the extraction stage was superior to the minopool in the reverse tanscription stage for detection of HCV cDNA.

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