Development and application of a SYBR Green I fuorescent PCR assay for the diferentiation of genotypes I and II African swine fever viruses  

作　　者：Xuexiang Yu Haowei Wu Dongfan Li Qian Xu Xiaoyu Chen Chengjun Zhang Fengqin Xu Xugang Ku Qigai He Wentao Li 

机构地区：[1]College of Veterinary Medicine,Huazhong Agricultural University,Wuhan,China [2]National Key Laboratory of Agricultural Microbiology,Wuhan,China [3]Hubei Hongshan Laboratory,Wuhan,China [4]The Cooperative Innovation Center for Sustainable Pig Production,Wuhan,China

出　　处：《Animal Diseases》2024年第4期289-295,共7页动物疾病（英文）

基　　金：supported by the National Key Research and Development Program of China(Grant No.2023YFF1000901);the Hubei Agricultural Research System(grant number HBHZD-ZB-2020-005);the National Key Research and Development Program of China(grant number 2021YFD1800101-2);Hubei Hongshan Laboratory(No.2022 hszd023);Project 2662023DKPY004;supported by the Fundamental Research Funds for the Central Universities.

摘　　要：African swine fever(ASF)is a highly fatal hemorrhagic disease afecting domestic pigs caused by African swine fever virus(ASFV).Genetic analysis of ASFV isolates to date has identifed 24 geographically related genotypes with various subgroups,but only genotype I and II ASFVs have been reported outside Africa.ASFV genotype II and genotype I viruses were reported in China in 2018 and 2021,respectively.In this study,unique and highly conserved noncoding regions were found between MGF_505-9R and MGF_505-10R in the 188 genomes of ASFV genotypes I and II.A pair of primers was designed on the basis of this region.By optimizing the reaction system and conditions,a SYBR Green I fuorescence PCR assay that can distinguish between ASFV genotypes I and II was established,and the sensitivity,reproducibility and specifcity were evaluated.The detection limit was 1 TCID_(50)/0.1 mL for both genotypes,with no cross-reactivity observed with other common pig pathogens.The intra-and interbatch variation coefcients were both less than 1.2%.Clinical sample detection analysis revealed 47 positive cases out of 100,including 3 for genotype I and 44 for genotype II,aligning with results from the WOAH-recommended and national standard methods.The method developed in this study allows for the diferentiation of ASFV genotypes I and II without the need for genome sequencing,ofering a convenient and rapid approach for ASFV detection and genotype identifcation.

关 键 词：African swine fever virus Genotype II Genotype I SYBR Green I fuorescent PCR DIAGNOSIS 

分 类 号：S852.65[农业科学—基础兽医学]