摘要
目的 探讨磁珠法提取核酸检测丙肝病毒RNA载量的方法学评价及其临床应用.方法 采用精密度、检测下限、线性分析和干扰实验对磁珠法提取核酸进行方法学评价,采用磁珠法和柱抽提法对283份标本进行检测,进行相关性分析.结果 磁珠法的高值质控血清的批内精密度为6.64±0.11,CV为1.65％,批间精密度为6.68±0.14,CV为2.09％;低值质控血清的批内精密度为4.60±0.12,CV为2.66％,批间精密度为4.56±0.19,CV为4.10％;检测下限为用磁珠法检测20份500IU/L HCV血清标本,20份标本均扩增出曲线,阳性率为100％（20/20）.磁珠法线性评估结果呈明显线性,相关系数r=0.999,P＜0.05,回归方程y=0.982x＋ 0.092.磁珠法和柱抽提法提取核酸检测HCV-RNA载量的比较,磁珠法与柱抽提法的相关系数r=0.990,P＜0.05,回归方程y=0.965x ＋0.247.结论 磁珠法提取核酸检测丙肝病毒RNA载量的方法具有较高的特异性和灵敏度,较强的抗干扰能力,且方法简便易于自动化操作,是理想的实验室核酸提取方法.
Objective To evaluate the magnetic beads-based nucleic acid extraction method for quantitative detection of hepatitis C virus （HCV）-RNA load and its clinical application.Methods The evaluation for the magnetic beads based nucleic acid extraction method was carried out including precision,limit of detection,linearity and anti-interference.The magnetic beads method was compared with column extraction technique to detect HCV-RNA load with 283 HCV serum samples.Results The high value control serum within run precision was 6.64 ± 0.11,CV was 1.65%,high value control serum between run precision was 6.68 ± 0.14,CV was 2.09%,the low value control serum within run precision was 4.60 ± 0.12,CV was 2.66%,low value control serum between run precision was 4.56 ± 0.19,CV was 4.10%.20 HCV serums was detected with magnetic beads method to evaluate limits of detection,all serums were amplified curve,positive rate was 100% （20/20）.Magnetic beads method of linear evaluation results showed that r =0.999,P 〈 0.05,regression equation was Y =0.982x ＋ 0.092.Comparison of magnetic beads method and column extraction technique to extract nucleic acid detection HCV-RNA load,there is a clear linear,r =0.999,P 〈 0.05,regression equation was Y =0.965x ＋ 0.247.Conclusion The magnetic beads-based nucleic acid extraction method has high specificity and sensitivity,strong anti-interference ability,and simple and easy for automation,it is an ideal laboratory nucleic acid extraction method.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2014年第6期482-484,共3页
Chinese Journal of Experimental and Clinical Virology
基金
国家临床重点专科建设项目（检验专业）资金资助
关键词
肝炎
丙型
聚合酶链反应
免疫磁化分离
RNA
Hepatitis C
RNA
Polymerase chain reaction
Immunomagnetic separntion
