柯萨奇病毒A16型核酸检测试剂国家参考品的建立

Establishment of national reference panel for coxsackievirus A16 nucleic acid detection kit

目的 建立柯萨奇病毒A16型(coxsackievirus A16,CA16)核酸检测试剂国家参考品,并制定其质量标准。方法收集并筛选CA16核酸阳性和阴性样本,分装、冻干后,建立CA16核酸检测试剂国家参考品。采用不同企业的试剂对参考品进行协作标定,根据协作标定结果确定参考品的质量标准,并考察其均匀性和稳定性。结果 CA16核酸检测试剂国家参考品由9份阳性参考品,8份阴性参考品,1份检测限参考品和1份精密度参考品组成。其质量标准为:阳性参考品符合率≥8/9;阴性参考品符合率为8/8;最低检测限为稀释度不低于1∶103;精密度参考品稀释100倍后重复检测10次,结果均为阳性,且Ct值的CV≤5%。该参考品均匀性符合要求,室温(25℃)放置1 d、反复冻融3次均未影响参考品的稳定性。结论 建立了首批CA16核酸检测试剂国家参考品,并制定了质量标准,为相关试剂的质量控制和评价提供了依据。

Objective To establish the national reference panel for coxsackievirus A16(CA16)nucleic acid detection kit and related quality standard.MethodsThe CA16 positive and negative samples were collected and screened,and then were filled and lyophilized to establish the national reference panel for CA16 nucleic acid detection kit.According to the cooperative calibration results of various reagent manufacturers,the quality standard of reference panel was determined.Meanwhile,the homogeneity and stability of the national reference panel were well studied.ResultsThe national reference panel of CA16 nucleic acid detection kit consisted of 9 positive samples,8 negative samples,1 limit-detecting sample and1 precision sample.The quality standard was as follows:the coincidence rate of positive samples was no less than 8/9;The coincidence rate of negative samples was 8/8;The minimum detection limit required that the dilution of limit-detecting sample was no less than 1∶103;The precision required that the coefficient of variation(CV)of Ct value of 10 precision samples diluted 100 times was no higher than 5% and the results were all positive.The homogeneity of the reference panel met the requirement,and the stability was not affected by the storage at room temperature(25 ℃)for 24 hours and repeated freezing and thawing three times.ConclusionThe first national reference panel of CA16 nucleic acid detection kit and the related quality standard have been established,which provided a reference for the quality control and evaluation of the related reagents.