中国畜牧兽医2017，Vol.44Issue(11)：3287-3293,7.DOI:10.16431/j.cnki.1671-7236.2017.11.025
含非洲猪瘟病毒核酸序列病毒样颗粒的研制及其在检测方法中的应用
Development of Virus-like Particles Containing African Swine Fever Virus Nucleic Acid Sequence and Its Application in Detection Method
摘要
Abstract
African swine fever (ASF) is an infectious disease caused by the African swine fever virus (ASFV).In order to ensure the accuracy and reliability of the test results,it is necessary to develop positive standard control products used in the kit.The study was aimed to develop the virus-like particles containing African swine fever virus (ASFV) nucleic acid sequence and its application in detection method.Fristly,the full-length gene fragment of p72 gene was amplified,and the ASF DNA virus-like particles containing p72 gene was constructed using insect baculovirus system.In order to further validate the reliability of the virus-like particles in the application of the method,DNA nucleic acid was extracted simultaneously with the cultured virus and infected tissue sample and applied in the Real-time quantitative PCR method.The results showed that the virus-like particles prepared by this study could replace the ASFV as a positive control product in the Real-time quantitative PCR method,and could act as quality control during nucleic acid extraction process.In the fluorescent PCR detection kit,the lowest packaging concentration of virus-like particles was 102 TCID50.Further studies had shown that the virus-like particles were also suitable for common PCR and LAMP detection methods,the minimum concentration were 103 and 101 TCID50,respectively.The results of this study would be important for the ASF detection method,promoting the application of the method and ensuring the accuracy and reliability of the test results.关键词
非洲猪瘟/参考物质/病毒样颗粒/检测方法Key words
African swine fever (ASF)/reference substance/virus-like particle/detection methods分类
农业科技引用本文复制引用
冯春燕,杜方原,刘丹丹,Pershin Andrey,王彩霞,张永宁,吴绍强,林祥梅..含非洲猪瘟病毒核酸序列病毒样颗粒的研制及其在检测方法中的应用[J].中国畜牧兽医,2017,44(11):3287-3293,7.基金项目
十三五科技支撑计划(2013BAD12B00) （2013BAD12B00）
国家质检总局科研基金资助项目(2015IK310) （2015IK310）
