贵州医科大学学报

2017, v.42;No.205(10) 1139-1144

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MAPREC技术检测Sabin株脊髓灰质炎病毒的基因突变率并评价疫苗神经毒力
MAPREC Assay to Detect the Gene Mutation Rate of Sabin Inactivated Poliovirus Vaccine and Evaluate Neurovirulence

姬光;平玲;王多义;沈武玲;王磊;寸怡娜;孙明波;
JI Guang;PING ling;WANG Duoyi;SHEN Wuling;WANG Lei;CUN Yina;SUN Mingbo;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases;

摘要(Abstract):

目的:采用MAPREC技术检测Sabin株脊髓灰质炎灭活疫苗(s IPV)生产收获液中病毒神经毒力决定位点的基因突变并评估疫苗神经毒力。方法:采用MAPREC技术用CY5荧光标记引物替代同位素标记检测疫苗株SabinⅠ、Ⅱ型及PfizerⅢ型疫苗病毒收获液及毒种的神经毒力决定位点的突变率,并与猴体神经毒力比较;采用MAPREC技术检测在不同培养温度、传代的次数、降低病毒接种的感染复数(MOI)时病毒各位点突变率变化。结果:MAPREC方法检测显示,本研究收集的病毒收获液的突变率符合猴体神经毒力检测结果;当病毒培养条件发生改变导致突变率增加后,MAPREC检测出的神经毒力决定位点的突变率也同样增加。结论:MAPREC技术可用于检测Sabin株脊髓灰质炎病毒的基因突变率,并评价疫苗的神经毒力。
Objective: Adopting the MAPREC to test mutation rate of Sabin IPV and the neurovirulence. Methods: The isotope labeling of MAPREC technique was replaced by CY5 fluorescent labeled primer,and detect Sabin vaccine strain type Ⅰ,Ⅱ,and Pfizer Ⅲ vaccine virus harvest and nerve toxicity of the poison of decision locus mutation rate; to test whether the results consistent with the monkeys nerve toxicity test results. When the temperature of virus culture and the number of viral passages was increased,multiplicity of infection was decreased,the mutation of virus would be increased correspondingly. Results: The rate of mutation detected by the qualified vaccine MAPREC method was consistent with the results of monkeys nerve toxicity test. When the virus culture conditions changed and the mutation rate increased,the mutation rate of the nerve virulence localization sites detected by MAPREC also increased. Conclusion: MAPREC test can detect the gene mutation rate of SIPV,so as to determine the vaccine's neurovirulence.

关键词(KeyWords): 脊髓灰质炎灭活疫苗;MAPREC实验;神经毒力决定位点;突变率;感染复数
inactivated poliomyelitis vaccine;MAPREC test;neurovirulence determinate sites;mutation ratio;multiplicty of infection

Abstract:

Keywords:

基金项目(Foundation): 中国医学科学院重大协调创新项目(2016-I2M-1-019);; 云南省重大科技专项(2016ZF003)

作者(Author): 姬光;平玲;王多义;沈武玲;王磊;寸怡娜;孙明波;
JI Guang;PING ling;WANG Duoyi;SHEN Wuling;WANG Lei;CUN Yina;SUN Mingbo;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases;

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DOI: 10.19367/j.cnki.1000-2707.2017.10.006

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