两种核酸提取方法检测丙肝HCV-RNA定量的比较

Comparison of two nucleic acid extraction methods in quantitative detection of HCV-RNA

目的:比较两种HCV-RNA核酸提取试剂对丙肝RNA病毒定量检测结果的差异,选出符合临床实验室要求的方法。方法:本实验从佛山市第一人民医院Roche全自动核酸分析已检测患者样本中选取浓度梯度分布均匀的阳性标本30例,分别使用中元磁珠法和达安磁珠法两种方法进行核酸提取,同时使用达安手工法进行平行实验,采用实时荧光定量PCR技术进行扩增和产物分析,利用统计学方法评价中元磁珠法和达安磁珠法两种核酸提取方法检测结果的正确度以及与达安手工法检测结果的一致性。结果:两种核酸提取方法检测丙肝HCV-RNA定量结果的正确度均可接受,中元磁珠法与达安手工法的一致性不及达安磁珠法与手工法的一致性好。结论:达安磁珠法正确度及与手工法的一致性更符合临床实验室的要求。

Objective:To compare the difference in quantitative detection of HCV-RNA using two different HCV-RNA nucleic acid extraction reagents,so as to identify the detection method that meets the requirements of clinical laboratory.Methods:In this experiment,30 positive specimens with evenly-distributed concentration gradient were selected from Roche full-automatic nucleic acid analysis system in Foshan First People’s Hospital.For each sample,nucleic acid extraction was performed using the Zhongyuan magnetic bead method and Daan magnetic bead method,respectively.Meanwhile,a parallel detection were performed using Daan manual method.Real-time quantitative PCR was used for amplification and product analysis.The detection accuracy rates with Zhongyuan and Daan magnetic beads and their consistency with Daan manual method were analyzed statistically.Results:The accuracy of the two nucleic acid extraction methods for quantitative detection of HCV-RNA in hepatitis C appeared acceptable.Compared:with Daan magnetic bead method,the findings of Zhongyuan magnetic bead method were less consistent with Daan manual method Conclusion The accuracy of Daan magnetic bead method and its consistency with the Daan manual method are more suitable for clinical laboratory practice.