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Establishment and Application of a SYBR Green I Real-time Quantitative PCR Assay for the Detection of LAT Gene of Pseudorabies Virus in Bama Miniature Pigs

查看全文 作　　者：Sufang [1]LU;Guangjun [1]GUO;Ling [1]MO;Feng [1]WEI;Bingmei [2]DONG;Wentong [2]ZHANG;Zhiqiang [1,2]SHEN 高影响力作者 机构地区：[1]Shandong Binzhou Animal Science & Veterinary Medicine Academy/Shandong Research & Development Center of Veterinary Propolis Vaccines;[2]Shandong Lvdu Bio-Sciences & Technology Co.,Ltd.高影响力机构 出　　处：《Agricultural Biotechnology》索引2015年第4卷第3期,共5页高影响力期刊 基　　金：Supported by Shandong Provincial Natural Science Foundation(ZR2012CQ012);Shandong Provincial Technical Innovation Grant(201220916006);Special Fund for Applied Technology Research and Development of Binzhou City(200706) 摘　　要：[Objective] This study aimed to establish a rapid,sensitive and specific SYBR Green I real-time quantitative PCR assay for the detection of latent pseudorabies virus( PRV) infection. [Method]SYBR Green I real-time quantitative PCR conditions and system for early detection of latent pseudorabies virus infection were optimized and compared with conventional PCR to investigate the sensitivity and specificity. Subsequently,the established assay was applied to detect different clinical samples. [Result]The sensitivity of SYBR Green I real-time quantitative PCR assay( 52 copies / μl) was 1 000 times higher than that of conventional PCR( 5. 2 × 104 copies / μl) and the detection time was shortened by 1 /2. The established assay could be used to detect PRV but could not be used to detect PCV2,PPV,CSFV or PRRSV. Various tissues were collected from Bama miniature pigs with latent PRV infection under sterile conditions for real-time PCR detection. Results showed that viral copy number in the brain,nasal swab,inguinal lymph node,liver,lung and spleen was above 20,while PRV was not detected in the kidney and heart tissues. [Conclusion]The established SYBR Green I real-time quantitative PCR assay for PRV LAT detection was specific,sensitive and rapid,which could be used for pathogen monitoring,epidemiological investigation and quantitative study of PRV. 关 键 词：实时荧光PCR检测 猪伪狂犬病病毒 SYBR 病毒基因 巴马小型猪 猪繁殖与呼吸综合征病毒 LAT 实时定量
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