谷金莲,于洋,梁争论
GU Jin-lian,YU Yang,LIANG Zheng-lun

• 1:中国食品药品检定研究院

摘要(Abstract)：

目的研制第6套丙型肝炎病毒(hepatitis C virus,HCV)抗体诊断试剂国家参考品。方法从我国10省市血站及生物制品企业的供血员中筛选候选参考品血浆样本304份,采用国内外27种检测试剂进行HCV Ag、HCV Ab及HCV RNA检测后,应用CHIRON公司的RIBA HCV 3.0 SIA及MP公司HCV BLOT 3.0试剂进行确证检测;利用Abbott Real Time HCV GenotypeⅡ试剂,对HCV RNA阳性样本进行HCV基因型别检测,并以核苷酸序列测定方法对分型样本进行确认。通过分析上述检测数据,从中拟选出协作标定样本,邀请国内外生产或市场使用占较大份额的11个实验室参加该套参考品标定。取最低检出限参考品4份,分别于4、25和-20℃放置1、3、5、7、9和11 d,或置-70℃反复冻融1~6次,进行稳定性试验。结果选出65份样品组成第6套HCV抗体诊断试剂国家参考品,其中包括阴性参考品30份,阳性参考品30份,最低检出限参考品4份,精密性参考品1份。应用11家的16种HCV抗体试剂分别平行3次标定检测,阴、阳性参考品符合率均≥29/30;最低检出限参考品检测L1~L3均为阳性,L4为阴性;精密性参考品检测CV在1%~11%之间。不同温度保存11 d及-70℃保存反复冻融6次的检测结果均符合要求(P>0.05)。结论建立了应用于血液筛查丙型肝炎病毒抗体试剂质量控制的第6套HCV抗体国家参考品。
Objective To prepare the 6th series of national reference for anti-hepatitis C virus(HCV)diagnostic reagent.Methods A total of 304 plasma samples of candidate references were screened and collected from healthy blood donors in blood centers and manufacturers of biological products in 10 provinces and or municipalities in China,in which HCV Ag,HCV Ab and HCV RNA were tested by 27 kinds of detection reagents at home and abroad,and confirmed by RIBA HCV 3.0 SIA from CHIRON and HCV BLOT 3.0 reagent from MP.The HCV genotypes of HCV RNA positive samples were identified by Abbott Real Time HCV Genotype Ⅱ reagent,and confirmed by nucleotide sequencing.By means of analyzing the above-mentioned test data,the samples for collaborative calibration by 11 laboratories occupying large shares in production or market at home and abroad.Four references for minimum detection limits were stored at 4,25 and-20 ℃for 1,3,5,7,9 and 11 d,or subjected to repeat freeze-thawing for 1 ~ 6 times after storage at-70 ℃ for stability test.Results A total of 65 samples are selected to compose the 6th series of national reference for anti-HCV diagnostic reagent,including 30 negative references,30 positive references,4 references for minimum detection limit and one reference for precision.The results of 3 parallel calibrations with 16 kinds of anti-HCV reagents from 11 laboratories showed that both the coincidence rates of negative and positive references were not less than 29/30.The test results of references L1 ~ L3 for minimum detection limits were positive,while that of L4 was negative.The CVs of test results of reference for precision were 1% ~ 11%.However,the test results of references after storage at various temperatures for11 d and after freeze-thawing for 6 times after storage at-70 ℃ met the relevenat requirements(P > 0.05).Conclusion The 6th series of national reference for anti-HCV diagnostic reagent used for quality control of blood screening of antiHCV reagent was developed.

关键词(KeyWords)： 丙型肝炎病毒;抗体;诊断试剂;参考品
Hepatitis C virus(HCV);Antibody;Diagnostic reagent;Reference

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 谷金莲,于洋,梁争论
GU Jin-lian,YU Yang,LIANG Zheng-lun

DOI: 10.13200/j.cnki.cjb.001374

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