献血员丙肝抗体筛查试剂盒的使用情况及质量控制
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摘要
丙型肝炎是1989年在日本国际肝炎学术会议上正式命名的一种新型肝炎。丙型肝炎病毒(Hepatitis C Virus，HCV)是导致输血后肝炎的一个重要病因。流行病学调查表明，全世界输血后肝炎的发生率平均约为1％，其中98％为丙型肝炎。丙肝主要的传播途径是输血和注射，抗-HCV阳性的献血员是传播丙肝的最危险的传染源。因此，严格把好献血员丙肝病毒感染检测的质量关，是切断输血传播丙肝、保证原料血浆及临床用血安全性的关键。
目前，国内大、中、小型采供血服务部门主要采用酶联免疫吸附试验(Enzyma Linked Immunnosorbent Assay，ELISA)检测丙型肝炎病毒抗体，以此作为筛选供血者资格的指标之一。自1994年我国卫生部规定对献血员血样检验体外免疫诊断试剂实施鉴定以来，能通过鉴定的抗-HCV ELISA试剂盒众多。近年来研究表明：各种试剂之间在灵敏度和特异性存在着差异。本课题的目的是：(1)调查近两年来我单位常用的几种抗-HCV ELISA试剂盒的使用情况，(2)采用实验手段，制
备出能对试剂进行质量评价的一套质控血清，作为我们今后
工作中选择试剂盒的参考依据。
本课题首先对2001年、2002年两年来我单位使用不同
厂家抗一HCV ELISA试剂盒对献血员丙肝抗体筛检的情况进
行统计分析;然后，收集2002年3月至12月献血员丙肝抗
体筛检阳性的原料血浆(100或200ML)及相应的检测标本
120例，一20℃以下冰冻保存，随机选择其中的100例标本，
用我们常用的四种ELISA试剂对标本进行检测分析;第三，
在这100例标本中，随机选只有一种ELISA试剂筛查阳性的
标本20例做HCVRNA检测，随机选两种及两种以上ELISA
试剂筛查阳性的标本60例做HCVRNA检测;第四，对做
RT一PCR的所有标本，采用HCV抗体分片段酶联免疫确认试
剂进行分片段检测;第五，对各种不同片段阳性的原料血浆
进行病毒灭活、分装，以作质控品用。
本课题结果表明，常用的四种厂家的抗一HCV检测试剂盒
的检测结果存在显著差异;ELISA法与PCR法检测丙型肝炎
结果也存在差异;分片段酶联免疫确认试剂能从HCV及
HCVRNA检测阳性标本中检测出不同阳性片段组合的标
2
本。
结论，目前市场上各种厂家生产的抗一HCV试剂盒在灵
敏度、特异性上存在着差异，因此，用户在使用时一定要谨
慎选择。采用HCV抗体分片段酶联免疫确认试剂进行分片
段检测
分装、
得到了各种不同片段阳性组合的血浆，经过灭活、
保存制备成质控品，以用于今后工作中挑选抗一HCV
酶联免疫试剂盒的参考依据。
Hepatitis C is a new tapy hepatitis named by internation hepatitis academic conference in Japan in 1989 . Hepatitis C Vims (HCV) is a most reasion casing transfused hepatitis. Epidemiology reseach indicates that transfused hepatitis incidence rate is 1% average in the word,and among the total 98% are hepatitis C.Hepatitis C primary route of transmission is transfiision and injection.Anti-HCV positive blood donors are the most dangerous soure of infection to transmit hepatitis C. Therefore rigorous HCV measure is the key to cut off the transmission of hepatitis C and to assure plasma and blood security.
The Mnistry of Health has brought vitro immunosorbent diagnosis reagent identify to blood donor into effect in 1994 and lots of anti-HCV kit can through the identify. Reseach indicates that there are different sensitivity and specificity among sorts of
kits.The purpose of this article are:(l) to reseach the use of some kits we use in recent years; (2) in order to progress reagent quality evaluate in our routine work we prepare a set of quality serum as the reference to choose the kits .
First ,this article analyzy different conditions of anti-HCV kits statistical in 2001 and 2002;second,to collect 120 sample of anti-HCV positive plasmar or blood from March to December in 2002,freezing conservation at -20,random choose 100 samples to analyze using four kinds ELISA kits we usually used;third,random choosing 80 sample from the 100 samples detect their HCVRNA;forth ,all 80 samples detect them using anti-HCV different region enzyma linked immunosorbent kit;fifth,virus inactivation and part pack for kinds of positive plasma.
The result indicate that there are differences among the four kinds of anti-HCV kits;that there are difference between ELISA and PCR to detect HCV;that anti-HCV different region enzyma linked immunosorbent kit can deride diferent combined segment
enzyma sample.
Now there are great differences at sensitivity and specificity in difference ant-HCV kit.SO the consumers must be cautious to choose ant-HCV kit.To detect samples using Anti-HCV different Region ELISA Kit we can receive sorts of different segments positive combination plasma,inactivation and part pack to conserve and prepare quality control serum as the reference to choose the kits.
目前，国内大、中、小型采供血服务部门主要采用酶联免疫吸附试验(Enzyma Linked Immunnosorbent Assay，ELISA)检测丙型肝炎病毒抗体，以此作为筛选供血者资格的指标之一。自1994年我国卫生部规定对献血员血样检验体外免疫诊断试剂实施鉴定以来，能通过鉴定的抗-HCV ELISA试剂盒众多。近年来研究表明：各种试剂之间在灵敏度和特异性存在着差异。本课题的目的是：(1)调查近两年来我单位常用的几种抗-HCV ELISA试剂盒的使用情况，(2)采用实验手段，制
备出能对试剂进行质量评价的一套质控血清，作为我们今后
工作中选择试剂盒的参考依据。
本课题首先对2001年、2002年两年来我单位使用不同
厂家抗一HCV ELISA试剂盒对献血员丙肝抗体筛检的情况进
行统计分析;然后，收集2002年3月至12月献血员丙肝抗
体筛检阳性的原料血浆(100或200ML)及相应的检测标本
120例，一20℃以下冰冻保存，随机选择其中的100例标本，
用我们常用的四种ELISA试剂对标本进行检测分析;第三，
在这100例标本中，随机选只有一种ELISA试剂筛查阳性的
标本20例做HCVRNA检测，随机选两种及两种以上ELISA
试剂筛查阳性的标本60例做HCVRNA检测;第四，对做
RT一PCR的所有标本，采用HCV抗体分片段酶联免疫确认试
剂进行分片段检测;第五，对各种不同片段阳性的原料血浆
进行病毒灭活、分装，以作质控品用。
本课题结果表明，常用的四种厂家的抗一HCV检测试剂盒
的检测结果存在显著差异;ELISA法与PCR法检测丙型肝炎
结果也存在差异;分片段酶联免疫确认试剂能从HCV及
HCVRNA检测阳性标本中检测出不同阳性片段组合的标
2
本。
结论，目前市场上各种厂家生产的抗一HCV试剂盒在灵
敏度、特异性上存在着差异，因此，用户在使用时一定要谨
慎选择。采用HCV抗体分片段酶联免疫确认试剂进行分片
段检测
分装、
得到了各种不同片段阳性组合的血浆，经过灭活、
保存制备成质控品，以用于今后工作中挑选抗一HCV
酶联免疫试剂盒的参考依据。
Hepatitis C is a new tapy hepatitis named by internation hepatitis academic conference in Japan in 1989 . Hepatitis C Vims (HCV) is a most reasion casing transfused hepatitis. Epidemiology reseach indicates that transfused hepatitis incidence rate is 1% average in the word,and among the total 98% are hepatitis C.Hepatitis C primary route of transmission is transfiision and injection.Anti-HCV positive blood donors are the most dangerous soure of infection to transmit hepatitis C. Therefore rigorous HCV measure is the key to cut off the transmission of hepatitis C and to assure plasma and blood security.
The Mnistry of Health has brought vitro immunosorbent diagnosis reagent identify to blood donor into effect in 1994 and lots of anti-HCV kit can through the identify. Reseach indicates that there are different sensitivity and specificity among sorts of
kits.The purpose of this article are:(l) to reseach the use of some kits we use in recent years; (2) in order to progress reagent quality evaluate in our routine work we prepare a set of quality serum as the reference to choose the kits .
First ,this article analyzy different conditions of anti-HCV kits statistical in 2001 and 2002;second,to collect 120 sample of anti-HCV positive plasmar or blood from March to December in 2002,freezing conservation at -20,random choose 100 samples to analyze using four kinds ELISA kits we usually used;third,random choosing 80 sample from the 100 samples detect their HCVRNA;forth ,all 80 samples detect them using anti-HCV different region enzyma linked immunosorbent kit;fifth,virus inactivation and part pack for kinds of positive plasma.
The result indicate that there are differences among the four kinds of anti-HCV kits;that there are difference between ELISA and PCR to detect HCV;that anti-HCV different region enzyma linked immunosorbent kit can deride diferent combined segment
enzyma sample.
Now there are great differences at sensitivity and specificity in difference ant-HCV kit.SO the consumers must be cautious to choose ant-HCV kit.To detect samples using Anti-HCV different Region ELISA Kit we can receive sorts of different segments positive combination plasma,inactivation and part pack to conserve and prepare quality control serum as the reference to choose the kits.
引文
[1] Takeuchi K,et al.Hepatitis C viral cDNA clones isolated from ahealthy carrier donor implicated in past-transfution non-A,non-B hepatitis,Gene. 1990,91(2):287.
[2] Alter H J.New kit on the block: evaluation of secondgeneration assays for detection of antibody to the hepatitis C virus.Hepatology 1992,15(2):350.
[3] Uytendaele S,et al.Evaluation of third-generation screening and confirmatory assays for HCV antibodys.Vox Ssng. 1994,66 (2):122
[4] 刘素芳．国家批批检定后抗．HCV诊断试剂的质量评价．中国输血杂志，1997．10(1)：36．
[5] Choo ql,et al.Brit Med Bulletin. 1990,46(2):423.
[6] 郝飞，余宙耀，主编．丙型肝炎基础与临床．北京：人民卫生出版社．1998，9．
[7] Benvegnu L,et al.Hepatology. 1997, 12(7):522-527.
[8] 梅志强，主编．人类新传染病概要．太原：山西科学技术出版社．2002，188-199．
[9]同[4]．
[10]陈均，等，多厂家ELISA试剂检测抗-HCV结果不一致的确认．中国输血杂志．2001，9(3)：26．
[11]刑文革，郑怀竞，等，国产丙型肝炎病毒抗体酶免检测试剂盒的质量评价．中华肝脏病杂志．2001，10(9)：314。
[12]Okamoto H,Kobata S,Tokita H, et al.Characterization of genomes sequence of tapy Ⅱ (or3a) hepatitis C virus isolates and PCR primers for specific detection.J.Gen.Virol.1993,74:2385-2390.
[13] Okamoto H,Kobata S,Sakamoto M,et al.A second-generation method of genotyping hepatitis C virus by the polymerase chain reaction with sense and antisense primers deduced from the core gene.J. Virol.Meth, 1996,57:31-45.
[14]同[8]．
[15]郝飞，余宙耀，主编．丙型肝炎基础与临床．北京：人民卫生出版社．1998．
[16]Berget A,Brener J,Doerr H,et al.Quantification of viral load: clinical relevance for human immunodeficiency virus
hepatitis B virus and hepatitis C virus infection [J].Intervirology. 1998,41(1):24-34
[17]肖星甫，主编．输血技术手册．成都：四川科学技术出版社，1992：226．
[18]张贺秋，等．丙型肝炎病毒不同区抗原酶免疫试剂研究．中华医学检验杂志．1999；22(6)：1-3．
[2] Alter H J.New kit on the block: evaluation of secondgeneration assays for detection of antibody to the hepatitis C virus.Hepatology 1992,15(2):350.
[3] Uytendaele S,et al.Evaluation of third-generation screening and confirmatory assays for HCV antibodys.Vox Ssng. 1994,66 (2):122
[4] 刘素芳．国家批批检定后抗．HCV诊断试剂的质量评价．中国输血杂志，1997．10(1)：36．
[5] Choo ql,et al.Brit Med Bulletin. 1990,46(2):423.
[6] 郝飞，余宙耀，主编．丙型肝炎基础与临床．北京：人民卫生出版社．1998，9．
[7] Benvegnu L,et al.Hepatology. 1997, 12(7):522-527.
[8] 梅志强，主编．人类新传染病概要．太原：山西科学技术出版社．2002，188-199．
[9]同[4]．
[10]陈均，等，多厂家ELISA试剂检测抗-HCV结果不一致的确认．中国输血杂志．2001，9(3)：26．
[11]刑文革，郑怀竞，等，国产丙型肝炎病毒抗体酶免检测试剂盒的质量评价．中华肝脏病杂志．2001，10(9)：314。
[12]Okamoto H,Kobata S,Tokita H, et al.Characterization of genomes sequence of tapy Ⅱ (or3a) hepatitis C virus isolates and PCR primers for specific detection.J.Gen.Virol.1993,74:2385-2390.
[13] Okamoto H,Kobata S,Sakamoto M,et al.A second-generation method of genotyping hepatitis C virus by the polymerase chain reaction with sense and antisense primers deduced from the core gene.J. Virol.Meth, 1996,57:31-45.
[14]同[8]．
[15]郝飞，余宙耀，主编．丙型肝炎基础与临床．北京：人民卫生出版社．1998．
[16]Berget A,Brener J,Doerr H,et al.Quantification of viral load: clinical relevance for human immunodeficiency virus
hepatitis B virus and hepatitis C virus infection [J].Intervirology. 1998,41(1):24-34
[17]肖星甫，主编．输血技术手册．成都：四川科学技术出版社，1992：226．
[18]张贺秋，等．丙型肝炎病毒不同区抗原酶免疫试剂研究．中华医学检验杂志．1999；22(6)：1-3．
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