不同厂家非洲猪瘟病毒荧光PCR检测试剂盒比对
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S851.3

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广东省重点领域研发计划资助项目(2019B020211005)


Comparison of Fluorescent PCR Kits Produced by Different Manufacturers for ASFV
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    摘要:

    为评价不同厂家非洲猪瘟病毒荧光PCR检测试剂盒的效果,选择当前市场上6个厂家的商品化试剂盒,检测不同类型样品,结合统计学方法对试剂盒的特异性、敏感性和重复性等性能指标进行比对分析。结果显示:6个厂家的试剂盒ROC曲线下区域面积(AUC)为0.826~0.951,Youden指数为0.76~0.90,说明试剂盒的排他性和包含性等特异性指标总体表现较好;最低检出限(LOD)等敏感性数据显示,6个厂家试剂盒的LOD为10-1~102 copies/μL,说明各试剂盒的敏感性均较理想;用高、低浓度样品对试剂盒分别进行批内、批间重复性试验,发现高浓度样品的批内、批间变异系数(CV)为0.10%~1.28%,说明重复性较好,仅有2个厂家的试剂盒在检测低浓度样品时表现较好,批内CV为2.25%~6.12%,批间CV为2.93%和4.93%,说明不同试剂盒检测低浓度样品时稳定性较差。本研究为商品化荧光PCR检测试剂盒的评价提供了参考。

    Abstract:

    In order to evaluate the effect of fluorescent PCR kits for African swine fever virus(ASFV)produced by different manufacturers,the kits supplied to current markets by six manufacturers were selected to detect different types of samples,then their performance indexes such as specificity,sensitivity and repeatability were compared and analyzed by statistical methods. The results showed that the area under ROC curve(AUC)of the kits ranged from 0.826 to 0.951,and the Youden index ranged from 0.76 to 0.90,indicating good performance of the exclusivity and inclusion in general;their LOD value ranged from 10-1 to 102 copies/μL,with good sensitivity. Based on the intra and inter batch repeatability tests with high and low concentration samples respectively,it was found that the coefficient of variation(CV)of high concentration samples was from 0.10% to 1.28%,with good repeatability. Good performance for samples with low concentration was observed in the kits produced by two manufacturers,the CV was from 2.25% to 6.12% for intra batch,2.93% and 4.93% for inter batch respectively,showing a poor stability when detecting samples with low concentration by different kits. A reference was thus provided for evaluation of commercial fluorescent PCR kits.

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许秀琼,刘劼,吴立炀,叶健,查云峰,王福广,孙柏林,卢受昇.不同厂家非洲猪瘟病毒荧光PCR检测试剂盒比对[J].《中国动物检疫》编辑部,2022,39(7):97-102.

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