两款荧光定量PCR仪检测HLA-B27基因结果比对分析

Comparative evaluation of HLA-B27 results detected by two different fluorescent quantitative PCR instruments

目的:探讨试剂与仪器匹配因素对PCR荧光法检测HLA-B27等位基因的影响。方法:首先采用自建荧光定量PCR法对85例样本中HLA-B27等位基因进行检测,随后采用商品化试剂盒PCR荧光法同时在两款荧光定量PCR仪Stratagene Mx3000P~?和Roche cobas~? Z480上对HLA-B27进行再次检测。结果:商品化试剂盒方法在cobas~? Z480上检出HLA-B27等位基因的结果为65例阳性、20例阴性,与自建方法所检出的结果完全一致;而其在Mx3000P~?上检出的结果为30例阳性、55例阴性,与自建方法的检测结果之间存在显著性差异(χ~2=33.03,P<0.005)。该商品化试剂盒方法在Mx3000P~?上的漏检率为53.85%。结论:该商品化试剂与Mx3000P~?荧光PCR仪的匹配性不佳,严重影响了检测结果的准确性。在临床实际工作中,评价PCR试剂的检测性能需要结合仪器,综合考虑以保证检测结果的准确性。

Objective:To explore the effect of reagent and instrument matching factors on the detection of HLA-B27 allele by real-time fluorescent PCR.Method:85 samples were detected for HLA-B27 allele by an in-house real-time PCR assay following by a commercial real-time PCR assay on two different fluorescent quantitative PCR instruments such as Stratagene Mx3000 P~?and Roche cobas~?Z480.Result:The results of the commercial assay on the Roche cobas~?Z480 were 65 positive and 20 negative.And they were identical to those detected by the in-house assay which was 65 positive and 20 negative.However,the results detected by the commercial assay on the Stratagene Mx3000 P~?were 30 positive and 55 negative,and they were different from the results detected by the in-house assay(χ~2=33.03,P<0.005).The missed rate of the commercial assay on the Stratagene Mx3000 P~?was 53.85%.Conclusion:The matching of the commercial reagent with Stratagene Mx3000 P~?is not well,which seriously affects the accuracy of the detection results.In clinical practical work,it is necessary to combine PCR reagents and instruments,comprehensively consider and evaluate detection performance to ensure the accuracy of results.