基于不同型别腺病毒标准品的呼吸道病原体核酸试剂最低检测限性能评价

Evaluation of the analytical sensitivity of twenty-eight respiratory pathogen nucleic acid detection kits based on different types of human adenovirus

目的利用不同型别呼吸道腺病毒核酸标准品对呼吸道病原体核酸检测试剂进行最低检测限性能评价。方法选择已研制的1、2、3、4、5、7、55型呼吸道腺病毒核酸标准品,参照美国临床实验室标准化协会(CLSI)的EP17-A2文件,采用Probit回归方法对28种不同方法学原理的呼吸道病原体核酸检测试剂的最低检测限(Lod)进行评估,比较各试剂盒声称Lod和评估Lod之间以及对不同腺病毒型别检测Lod之间的差异。结果28种试剂盒中能够准确评估Lod的比例为50%(12/24)。12种试剂盒的评估Lod与声称Lod一致性较好;12种试剂盒一致性较差,其中4种试剂盒差异较小(measured Lod<5×claimed Lod),8种试剂盒差异较大(measured Lod>5×claimed Lod),最大者可达24000倍。不同试剂间评估的Lod浓度差异可达106数量级以上(最低和最高分别为20拷贝/ml和4.8×10^(7)拷贝/ml);同种试剂对不同型别腺病毒的检测Lod均不相同,其浓度差异最高可达104数量级(最低和最高分别为5.0×10^(3)拷贝/ml和4.8×10^(7)拷贝/ml)。结论部分呼吸道病原体核酸检测试剂检测腺病毒的最低检测限性能评估缺乏准确性,使用检测范围内的多种病原体型别或亚型进行性能评估尤为必要。

Objective To evaluate the analytical sensitivity of twenty-eight respiratory pathogen nucleic acid detection kits based on different types of human adenovirus.Methods According to the EP17-A2 document of the Clinical and Laboratory Standards Institute,probit regression analysis of 95%hit rates was performed to evaluate the limit of detection(Lod)of 28 respiratory pathogen nucleic acid detection kits.The differences between claimed Lod and measured Lod,and between different adenovirus types were compared.Results The data showed that the claimed Lod for 50%(12/24)of the manufacturers can be accurately evaluated.There was a poor consistency in 12 kits,and the difference in 4 kits was not significant(measured Lod<5×claimed Lod),however,the difference in the other 8 kits was significant(measured Lod>5×claimed Lod,24000-fold increase for the most significant).The difference of measured Lod concentration between different kits was more than 106 orders of magnitude(the lowest and the highest were 20 copies/ml and 4.8×10^(7)copies/ml,respectively).An analysis of different types of adenovirus detected by the same kit revealed a wide range of measured Lod,spanning 104 orders of magnitude differences(the lowest and the highest were 5.0×10^(3)copies/ml and 4.8×10^(7)copies/ml,respectively).Conclusions It is showed that there is a lack of accuracy in evaluation of the analytical sensitivity for some respiratory pathogen nucleic acid detection kits,so it is particularly necessary to use a variety of pathogen types or subtypes for performance evaluation.