HCV RNA定量检测试剂的临床应用

The clinical application research of a reagent for quantification of HCV RNA

目的:探讨HCV RNA定量检测试剂的临床应用效果。方法:通过和市场上广泛使用的柱提取试剂比实验,对一种丙型肝炎病毒磁珠法核酸提取荧光定量PCR检测试剂的临床使用进行评估。结果:①浓度在5.00E+03～5.00E+06 IU/mL的样本,两种试剂的定量结果与理论靶值均具有很好的对应性,具有较好的重复性和精密度;对于1.00E+03IU/mL以下的样本,磁珠法的定量结果重复性好,定量准确;而柱提取法大部分均未能检出。②对于2种方法检测的339例临床样本,中高病毒载量组柱提取法检测>5.00E+02IU/mL有147例,两种试剂定量结果基本一致,相关性r为0.9413。3 339例临床血清样本中5例样本磁珠法检测为阳性,而柱提取法检测为阴性,P<0.01,二者有显著性差异。这5例样本,丙肝肝炎抗体检测均为阳性。结论:①对于中高病毒载量的样本,两种试剂均具有良好的检测效果;②磁珠法对比柱提取法在低病毒载量区域,灵敏度更高,同时操作相对简单,不易出现污染与脱靶,具有较高的临床应用价值。

The goal of this study is to evaluate a HCV RNA Quantitative Fluorescence Diagnostic Kit(refer to as magnetic beads method hereafter) in a clinic lab by comparing it with another kind of fluorescence diagnostic kit using column extraction technique,which is currently the most widely used method for nucleic acid quantitative test in China.The results of the serial experiment showed that: 1).The concentration range of 5.00E+03 to 5.00E+06IU/ml,the average concentration values from the two HCV RNA quantitative kits agree with each other and with the theoretical values very well.However,for samples with low concentration(<1.00E+03IU/ml),the magnetic beads method kit showed much better reproducibility,accuracy and sensitivity,while column extraction method kit failed to test positive for most of the low concentration samples.2).Among the 339 clinical samples,results from the two kits demonstrated that there was good agreement between the two reagents for the high viral load group(147 samples,column extraction method >500IU/mL),with a correlation r value of 0.941.3.5 conflicting samples which the magnetic beads method kit showed positive results,However the column extraction method kit failed to test positive of these samples,P<0.01,suggesting sensitivity difference between the two kits.5samples had HCV quantitative immunoassay results,and they showed good agreement with the results of the magnetic beads method kit.The results imply that 1.For samples in the medium and high concentration range,the two kits both showed good accuracy;2.The magnetic beads method showed much better accuracy and sensitivity than the column extraction method for the panel with low viral load.With a simple procedure,great accuracy and good sensitivity,this test kit is suitable for routine clinical lab usage.