不同保存条件对血清荧光定量HCV RNA检测的影响

Impact of various storage conditions on the detection of serum HCV RNA using fluorescence quantitation PCR

目的了解不同的保存条件对血清标本HCVRNA检测结果的影响。方法实时定量荧光聚合酶链法(FQ-PCR)检测已确诊的HCV患者30例。结果血清标本-20℃存放7d HCV RNA检测结果、加RNA酶抑制剂-20℃保存7d及-20℃反复冻融3次的HCV RNA检测结果差异无统计学意义。结论血清标本-20℃保存7d,加RNA酶抑制剂-20℃保存7d,-20℃反复冻融3次,不影响HCV RNA的检测结果。HCV RNA检测结果重复性不好,应该考虑内、外源性等RNase的降解作用及所用试剂的特异性、敏感性、重复性等因素的影响。

Objective To investigate the impact of various storage conditions of serum samples on HCV RNA detection.Methods Real-time fluorenscent quantitative PCR detected 30 examples have been diagnosed in patients with HCV.Results The results of HCV RNA in serum samples,which stored 7 days at -20℃,reserved 7 days with RNA enzyme inhibitor at -20℃ and repeated freezed-thawing three times respectively,showed no significant differences.Conclusion storage at -20℃ for 7 days,stourage for 7 days with RNA enzyme inhibitor and freezed-thawing three times repeatedly at -20℃ have no influence on detective result of HCV RNA in serum.That the repeatability is poor in detecting HCV RNA should consider the endogenous and exogenous degradation of RNase,and the effects of those factors such as specific,sensitivity and repeatability of the factors which were used.