三种HCV RNA荧光定量PCR检测试剂的结果分析及应用比较

Comparison of application of three types of HCV-RNA fluorescent quantitative PCR detection reagents

目的：比较3种丙型肝炎病毒（HCV）RNA荧光定量PCR检测试剂的临床应用性能。方法采用国产 A 试剂（磁珠分离法）、国产 B 试剂（荧光探针法）和目前国际临床广泛应用的 C试剂（内标法）同时检测110例丙型肝炎患者临床血液样本及梯度稀释的强阳性标本，从试剂间的相关性、检测灵敏度、特异度等对结果进行对比分析。结果A 和 B、A 和 C、B 和 C 试剂间的相关系数分别为0．845、0．917、0．860（P 均＜0．01）。在3种方法均有数值的58例标本中，3种试剂所检出的 HCV RNA 水平比较差异无统计学意义（P ＞0．05）。但在低病毒载量（＜103 IU ／ml）组中，3种试剂灵敏度比较差异有统计学意义（P ＜0．01），C 试剂灵敏度最高，A 试剂次之，B 试剂最低。当病毒载量在2．00＆#215;103～2．00＆#215;106 IU ／ml 时，3种试剂的定量结果与理论值均有很好的相关性与重复性，其检测浓度的平均值与理论浓度的双对数线性相关系数分别为0．9983、0．9823、0．9999（P 均＜0．01）；当病毒载量为2．00＆#215;102 IU ／ml 时，C 试剂的检测结果最接近，A 试剂次之，B 试剂未能检出。结论C 试剂作为国际上广泛应用于临床 HCV RNA 定量检测的试剂，优势明显；国产 A 试剂总体性能优于 B 试剂，且费用较国外试剂低廉，性价比较高。

Objective To compare the clinical application performance of three types of hepatitis C virus （HCV）-RNA fluorescent quantitative PCR detection reagents.Methods Domestic A reagent （magnet-ic bead separation method）,domestic B reagent （fluorescence probe method）and C reagent currently widely used internationally （internal standard method）were adopted to detect 1 1 0 clinical blood in patients with Hep-atitis and strong-positive samples with gradient dilution.The correlation,sensitivity and specificity were statisti-cally compared among the results of three detection reagents.Results The correlation coefficient of three types of reagents was 0.845,0.91 7 and 0.860 （all P 〈0.01 ）.In 58 samples detectable by three methods,no sig-nificant difference was found in HCV RNA level among three kinds of reagents （P 〉0.05）.However,in the low viral load group （〈1 .00 ×1 0^3 IU /ml）,the sensitivity significantly differed among three types of reagents with highest sensitivity in C reagent,followed by A and B reagents.In the viral load group （2.00 ×10^3 ～2.00 ×1 0^6 IU /ml）,high levels of correlation and repeatability between the quantitative results and theoretical val-ues were observed in all reagents.The double logarithmic linear correlation coefficient of the mean and theoreti-cal concentration was 0.9983,0.9823 and 0.9999 （all P 〈0.01 ）.When the viral load was 2.00 ×1 0^2 IU /ml,the detected concentration was the most similar to the theoretical concentration in C reagent,followed by A reagent and undetected in B reagent.Conclusions C reagent was superior to A and B reagents in the HCV-RNA fluorescent quantitative PCR detection.Compared with B reagent,A reagent exhibited better perform-ance.A reagent was cheaper compared with C reagent.Thus,A reagent could be used as a cost-effective HCV-RNA fluorescent quantitative PCR detection.