应用RT-PCR-RFLP鉴别2014年新余市麻疹病毒感染病例
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摘要
目的引进一种区别麻疹病毒疫苗株与野生株的简单方法,用以检测新余市麻疹病毒感染病例标本。方法新余市8份麻疹病毒感染病例标本和1株S191麻疹疫苗株经逆转录-聚合酶链反应(RT-PCR)扩增病毒H基因部分片段,应用限制性内切酶片段长度多态性分析(RFLP)方法进行酶切分析。结果 S191麻疹疫苗株被AflII限制性酶酶切成2个片段,8份麻疹病毒感染病例标本均未酶切消化。结论新余8例麻疹病例全部为麻疹野病毒所感染;RT-PCR-RFLP方法简单、有效,非常适合条件受限的实验室使用。
Objective To introduce a simple method for distinguishing the vaccine strain from wild strains of measles virus,and detect samples of measles virus-infected cases in Xinyu. Methods Eight cases of measles virus-infected and one S191 measles vaccine strain in Xinyu were amplified by reverse transcription-polymerase chain reaction(RT-PCR) to obtain partial fragments of viral H gene,and restriction endonuclease fragment length polymorphism(RFLP) analysis was used for restriction analysis. Results The PCR product of S191 vaccine strain was digested into 2 fragments by AflII restriction enzyme,while eight caess of measles virus-infected were not digested. Conclusion All eight patients were infected by measles wild viruses in Xinyu.RT-PCR-RFLP is a simple and effective method and it is very suitable for laboratories with limited conditions.
Objective To introduce a simple method for distinguishing the vaccine strain from wild strains of measles virus,and detect samples of measles virus-infected cases in Xinyu. Methods Eight cases of measles virus-infected and one S191 measles vaccine strain in Xinyu were amplified by reverse transcription-polymerase chain reaction(RT-PCR) to obtain partial fragments of viral H gene,and restriction endonuclease fragment length polymorphism(RFLP) analysis was used for restriction analysis. Results The PCR product of S191 vaccine strain was digested into 2 fragments by AflII restriction enzyme,while eight caess of measles virus-infected were not digested. Conclusion All eight patients were infected by measles wild viruses in Xinyu.RT-PCR-RFLP is a simple and effective method and it is very suitable for laboratories with limited conditions.
引文
[1]WHO. WHO/UNICEF Joint Statement:global plan for reducing measles mortality 2006-2010[EB/OL].[2018-06-19].http://www.unicef.org/media/files/globalplan.pdf.
[2]WHO. Global control and regional elimination of measles,2000-2012[J]. Wkly Epidemiol Rec,2014,89(6):45-52.
[3]Ma C,AN ZJ,Hao LX,et al. Progress toward measles elimination in the People’s Republic of China,2000-2009[J]. J Infect Dis,2011,204(Suppl17):447-454.
[4]马超,郝利斯,马静,等.中国2010年麻疹流行病学特征与消除进展[J].中国疫苗和免疫,2011,17(3):242-248.
[5]马超,苏琪茹,郝利斯,等.中国2012-2013年麻疹流行病学特征与消除麻疹进展[J].中国疫苗和免疫,2014,20(3):193-199.
[6]戴丽芳,唐宁,朱青,等.贵州省2008-2011年接种含麻疹成分疫苗后发热出疹性病例的流行病学分析[J].疾病监测与控制杂志,2013,7(10):585-587.
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[8]张双宅,史艳,蒋瑞平,等.石家庄市2013-2016年含麻疹成分疫苗接种偶合麻疹野病毒感染病例分析[J].中国疫苗和免疫,2017,23(5):526-529.
[9]周剑惠,王爽,陈超,等.中国麻疹病毒疫苗株与野毒株鉴别方法的建立[J].中国疫苗和免疫,2009,15(4):310-315.
[10]龚甜,熊英,周顺德,等.江西省2011年麻疹病毒野毒株与疫苗株的快速鉴别[J].现代预防医学,2013,40(6):3110-3112.
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[13]刁连东.麻疹[M].上海:上海科学技术出版社,2001:220-222.
[14]Duclos P,Ward BJ. Measles vaccines:a review of adverse events[J]. Drug Saf,1998,19(6):435-454.
[15]王真行,徐冰. WHO关于麻疹疫苗的意见书[J].国外医学:预防、诊断、治疗用生物制品分册,2005,28(1):8-12.
[16]Stanley AP,Walter AO,Paul AO.疫苗学[M].梁晓峰,译. 5版.北京:人民卫生出版社,2011:410.
[17]国家药典委员会.中华人民共和国药典:三部[M]. 2010年版.北京:中国医药科技出版社,2010:140.
[18]徐爱强,许青,宋立志,等.接种麻疹疫苗后发热出疹性病例的流行病学调查[J].中华流行病学杂志,2004,25(5):417-420.
[19]Mori T. A simple method for genetic differentiation of the ALK-C vaccine strain from wild strains of measles virus[J]. Biologicals,1994,22(2):179-185.
[20]张红波,莫尚鹏,陈玉红,等.宜春市2015年麻疹病毒野病毒感染病例的快速鉴别[J].实验与检验医学,2017,35(2):199-201.
[2]WHO. Global control and regional elimination of measles,2000-2012[J]. Wkly Epidemiol Rec,2014,89(6):45-52.
[3]Ma C,AN ZJ,Hao LX,et al. Progress toward measles elimination in the People’s Republic of China,2000-2009[J]. J Infect Dis,2011,204(Suppl17):447-454.
[4]马超,郝利斯,马静,等.中国2010年麻疹流行病学特征与消除进展[J].中国疫苗和免疫,2011,17(3):242-248.
[5]马超,苏琪茹,郝利斯,等.中国2012-2013年麻疹流行病学特征与消除麻疹进展[J].中国疫苗和免疫,2014,20(3):193-199.
[6]戴丽芳,唐宁,朱青,等.贵州省2008-2011年接种含麻疹成分疫苗后发热出疹性病例的流行病学分析[J].疾病监测与控制杂志,2013,7(10):585-587.
[7]刘冷,郑焕英,王慧玲,等.广东省含麻疹成分疫苗后发热出疹性病例的血清学和病毒学检测[J].中国疫苗和免疫,2017,23(3):257-261.
[8]张双宅,史艳,蒋瑞平,等.石家庄市2013-2016年含麻疹成分疫苗接种偶合麻疹野病毒感染病例分析[J].中国疫苗和免疫,2017,23(5):526-529.
[9]周剑惠,王爽,陈超,等.中国麻疹病毒疫苗株与野毒株鉴别方法的建立[J].中国疫苗和免疫,2009,15(4):310-315.
[10]龚甜,熊英,周顺德,等.江西省2011年麻疹病毒野毒株与疫苗株的快速鉴别[J].现代预防医学,2013,40(6):3110-3112.
[11]中华人民共和国国家卫生和计划生育委员会.中华人民共和国卫生行业标准:WS296-2017麻疹诊断[S].北京:[出版者不详],2017:12.
[12]王涛. 10例接种麻风疫苗后发热出疹性疾病的病例分析[J].现代预防医学,2011,38(6):1126-1127.
[13]刁连东.麻疹[M].上海:上海科学技术出版社,2001:220-222.
[14]Duclos P,Ward BJ. Measles vaccines:a review of adverse events[J]. Drug Saf,1998,19(6):435-454.
[15]王真行,徐冰. WHO关于麻疹疫苗的意见书[J].国外医学:预防、诊断、治疗用生物制品分册,2005,28(1):8-12.
[16]Stanley AP,Walter AO,Paul AO.疫苗学[M].梁晓峰,译. 5版.北京:人民卫生出版社,2011:410.
[17]国家药典委员会.中华人民共和国药典:三部[M]. 2010年版.北京:中国医药科技出版社,2010:140.
[18]徐爱强,许青,宋立志,等.接种麻疹疫苗后发热出疹性病例的流行病学调查[J].中华流行病学杂志,2004,25(5):417-420.
[19]Mori T. A simple method for genetic differentiation of the ALK-C vaccine strain from wild strains of measles virus[J]. Biologicals,1994,22(2):179-185.
[20]张红波,莫尚鹏,陈玉红,等.宜春市2015年麻疹病毒野病毒感染病例的快速鉴别[J].实验与检验医学,2017,35(2):199-201.