A群链球菌抗原检测试剂盒(胶体金法)质控参考品的建立及初步检测

Establishment and preliminary determination of quality control reference for evaluation of group A streptococcus antigen detection kit(immunochromatogragphic assay)

目的建立A群链球菌抗原检测试剂盒(胶体金法)质控参考品,并进行初步检测。方法从国家医学细菌保藏管理中心库存标准菌株中筛选出10株A群链球菌和10株非A群链球菌进行菌落形态、菌型、生物学特性及16s r RNA鉴定;并确定菌液浓度测定方法。结果选择了10株不同来源的A群链球菌作为阳性参考菌株,10株同属不同群或寄生部位与A群链球菌相同的其他种属菌株作为阴性参考菌株。经16s r RNA基因序列比对,所选参比菌株与相应的模式株相似度均在99%以上。平板计数结果明显低于显微计数法,不同人员分别对32份菌液进行显微计数的结果差异无统计学意义(P=0.310)。结论建立了A群链球菌抗原检测试剂盒(胶体金法)质控参考菌株,并确立了显微计数法作为确定参考菌液浓度的方法,为今后相关参考品的制备奠定了基础。

Objective To establish and preliminarily determine the quality control reference for evaluation of group A streptococcus antigen detection kit（immunochromatographic assay, ICA）. Methods Ten group A streptococcus strains and 10 non-group A streptococcus strains were screened out from National Center for Medical Culture Collections（CMCC） and identified for morphology, bacterial type, biological characters and 16 s rRNA, and the method for determination of concentration of bacterial liquid was screened. Results Ten group A streptococcus strains from various origins were selected as positive reference strains, while ten non-group A streptococcus strains sharing the same genus and parasitic site with group A streptococcus as negative reference strains. The comparison of 16 s RNA gene sequence exhibited the similarity values of more than 99% of all the selected strains to their closest type strains. The concentration of reference determined by plate counting method was significantly lower than that by microscopic counting method.However, the results of microscopic counting of 32 samples of bacterial liquids by various personnel showed no significant difference（P = 0. 310）. Conclusion The quality control reference strains for evaluation of group A streptococcus antigen detection kit were established, and microscopic counting method was selected for detection of concentration of reference,which laid a foundation of preparation of the relevant references.