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目的本研究拟在湖北省疾病预防控制中心对珠海海泰生物制药有限公司研制的麻疹、风疹病毒核酸荧光定量聚合酶链式反应(polymerase china reaction PCR)检测试剂盒开展临床试验,以对该试剂盒的各项性能进行评估。方法本试验设计采用单盲方法对我省范围内的134份咽拭子、尿液、漱口水、分离培养物样品进行检测,本研究中用海泰试剂和逆转录PCR(reverse transcription-PCR,RT-PCR)法对考核样品进行平行检测。采用国家下发的麻疹,风疹病毒分型引物分别进行RT-PCR,对RT-PCR产物进行凝胶电泳和基因测序分析。对与凝胶电泳结果不一致的样本,同时用海泰试剂和RT-PCR法重复试验并做凝胶电泳和基因测序分析。对于基因测序成功的样本,以基因测序法为准;基因测序失败的样本以凝胶电泳结果为准。最终结果以重复试验为准,通过对检测结果进行统计、分析,对海泰试剂的灵敏度、特异性、精密性及准确性等做出评价。结果在本试验中海泰试剂与参考方法相比较,其灵敏度为100%,特异度为92.9%,假阳性率为7.1%,假阴性率为0,粗一致性为99.3%,调整一致性为98%,约登指数为0.929,阳性预示值为99.2%,阴性预示值为100%。采用交叉四格表Kappa检验的统计方式进行分析,海泰试剂和参比试剂检测结果的一致率为99.3%,Kappa>0.75。结论评估试剂同参照试剂相比结果符合较好。该试剂盒为疾病的临床诊断、流行病调查提供了一种更为简捷精确,切实可行的检测手段。
OBJECTIVE: This study intends to carry out clinical trials of measles, rubella and nucleic acid fluorescence quantitative polymerase chain reaction (PCR) detection kits developed by Zhuhai Haitai Biological Pharmaceutical Co., Ltd. in Hubei Provincial Center for Disease Control and Prevention. The box’s performance is assessed. Methods In this study, 134 throat swabs, urine, mouthwash, and culture samples were tested by single blind method in our province. In this study, Haitai reagent and reverse transcription-PCR RT-PCR) method for parallel detection of samples tested. Measles and rubella virus typing primers were respectively used for RT-PCR and gel electrophoresis and gene sequencing analysis of RT-PCR products. Samples that were inconsistent with the results of gel electrophoresis were repeated with Haeimae reagent and RT-PCR and subjected to gel electrophoresis and gene sequencing analysis. For successful gene sequencing samples, the gene sequencing method shall prevail; the gene sequencing failure samples shall be subject to gel electrophoresis. The final result is based on repeated tests. The sensitivity, specificity, precision and accuracy of Haitai reagent are evaluated by statistical analysis of the test results. Results In this experiment, compared with the reference method, Haitai reagent has the sensitivity of 100%, the specificity of 92.9%, the false positive rate of 7.1%, the false negative rate of 0, the crude consistency of 99.3% and the consistency of adjustment of 98 %, Youden index was 0.929, the positive predictive value was 99.2%, the negative predictive value was 100%. Analyzed by the statistical method of Kappa test of crossover four grids, the coincidence rate of Haitai reagent and reference reagent was 99.3%, Kappa> 0.75. Conclusion The results of the assessment reagent are in good agreement with those of the reference reagent. The kit for the clinical diagnosis of disease, epidemiological investigation provides a more simple and accurate, practical detection means.