摘要
目的研制隐球菌荚膜多糖检测试剂国家参考品,并制定相关检测试剂的质量评价标准。方法收集不同型别隐球菌及相关菌株的临床分离株和标准菌株样本,采用飞行时间质谱法鉴定物种信息,有争议的样本通过宏基因组二代测序方法进行确证;对候选隐球菌阳性菌株中的标准菌株进行基因分型(PCR指纹法)及血清型(PCR法)鉴定。用筛选出的样本组成隐球菌荚膜多糖检测试剂国家参考品盘,并进行临床常用抗真菌药敏试验。多个厂家采用胶体金法或化学发光法隐球菌荚膜多糖检测试剂对国家参考品进行协作标定,确定相关检测试剂的质量标准,并检测其稳定性。结果共获得14份阳性参考品(P01~P14),10份阴性参考品(N01~N10),并以隐球菌标准菌株血清A型VNⅠ作为检出限参考品,隐球菌标准菌株血清AD型VNⅢ作为精密性参考品,组成隐球菌荚膜多糖检测试剂国家参考品盘。相关检测试剂的质量标准为:阴性参考品应至少9个样本为阴性,其他不作要求,阴性参考品符合率应不低于9/10;阳性参考品应均为阳性,符合率应为14/14;建议检出限参考品使用试剂盒样本处理液进行1∶200、1∶400、1∶800、1∶1600、1∶3200及1∶6400稀释(分别标记为S01~S06),S01~S03均应为阳性,其他不作要求;建议精密性参考品用试剂盒样本处理液进行1∶10及1∶100稀释,各平行检测10次,均应为阳性且显色度均一或10次检测结果变异系数(CV)≤15.0%。隐球菌荚膜多糖检测试剂国家参考品经25℃放置4 d及反复冻融5次均不影响该国家参考品的性能。结论研制的隐球菌荚膜多糖检测试剂国家参考品及制定的质量标准适用于相关检测试剂的质量评价需求。
Objective To develop the national reference panel for cryptococcal capsular polysaccharide antigen detection kit and establish the quality evaluation standards.Methods The clinical isolates and standard strain samples of different types of cryptococcus and related strains were collected and identified for the species information by time-of-flight mass spectrometry,and the controversial samples were confirmed by metagenomic second-generation sequencing.Genotyping(PCR fingerprinting)and serotype identification(PCR)were used to identify the standard strains of candidate cryptococcus positive strains.The selected samples were used as the national reference panel for cryptococcal capsular polysaccharide antigen detection kit,and the susceptibility test against commonly used clinical antifungal drugs was performed.Finally,the national reference panel was collaboratively calibrated by multiple manufacturers by using colloidal gold or chemiluminescence cryptococcal capsular polysaccharide antigen detection kit,the quality standards of the related detection reagents were determined,and the stability was evaluated.Results The reference panel consisted of 26 samples,including 14 positive reference materials(P01-P14),10 negative reference materials(N01-N10),1 precision reference material(VNⅢof cryptococcus standard strain serum type AD)and 1 limit of detection(LOD)reference material(VNI of cryptococcus standard strain serum type A).The quality standards were as follows:at least 9 of negative reference samples should be negative,others were not mandatory,and the coincidence rate of negative reference should be not less than 9/10;positive reference samples should all be positive,and the coincidence rate should be 14/14;it was suggested that the detection limit reference should be diluted at 1∶200,1∶400,1∶800,1∶1600,1∶3200 and 1∶6400 with the sample treatment solution of the kit(marked as S01-S06 respectively),of which S01-S03 should be positive and others were not required;it was suggested that the precision refe
作者
许庭莹
孙天舒
张丽
刘玲莉
井然
黄嘉维
张傲
许四宏
XU Tingying;SUN Tianshu;ZHANG Li;LIU Lingli;JING Ran;HUANG Jiawei;ZHANG Ao;XU Sihong(DivisionⅡof In Vitro Diagnostics for Infectious Diseases,Institute for In Vitro Diagnostic Reagents Control,National Institutes for Food and Drug Control,Beijing 100050,China;不详)
出处
《中国生物制品学杂志》
CAS
CSCD
2024年第11期1313-1318,1326,共7页
Chinese Journal of Biologicals
基金
国家传染病控制重大专项(2018ZX10102001)。
关键词
隐球菌
荚膜多糖
抗原检测试剂盒
国家参考品
Cryptococcus
Capsular polysaccharide
Antigen detection kit
National reference panel
