猫杯状病毒纳米PCR检测方法的建立  

Establishment of nano-PCR assay for detection of feline calicivirus

作　　者：赛力克·巴合达吾列提 刘存 刘砚涵 梁瑞英 秦彤[1,4] 梁琳 崔尚金[1,4] Sailike·bahedawulieti;LIU Cun;LIU Yan-han;LIANG Rui-ying;QIN Tong;LIANG Lin;CUI Shang-jin(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Shandong Provincial Center for Animal Disease Control,Jinan 250010,China;Altay Regional Center for Animal Disease Control and Prevention,Altay 836500,China;Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Technology of Beijing,Ministry of Agriculture,Beijing 100193,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]山东省动物疫病预防与控制中心,山东济南250010 [3]新疆阿勒泰地区动物疾病控制与诊断中心,新疆阿勒泰836500 [4]农业部兽用药物与兽医生物技术北京科学观测实验站,北京100193

出　　处：《中国兽医科学》2021年第6期684-688,共5页Chinese Veterinary Science

基　　金：中央级公益性科研院所基本科研业务费专项(2020-YWF-YTS-10);中国农业科学院创新工程项目(ASTIP-IAS15)。

摘　　要：为了提高猫杯状病毒(FCV)的检测效率,参考FCV全基因组序列的保守区域,利用生物学软件Primer Premier 5.0设计1对FCV特异性引物,并建立了FCV纳米PCR检测方法。结果显示,该方法特异性良好,与其他常见的猫病毒性病原均无交叉反应;该方法敏感性良好,最低检测量为5.4×10^(-3) pg/μL的标准品,是普通PCR的10倍。重复性试验结果显示,3批次重复试验结果保持一致,重复性良好。综上所述,建立的FCV纳米PCR方法具有良好的特异性、灵敏性和重复性,可作为FCV感染的实验室诊断的技术方法。In order to improve feline calicivirus(FCV)detection efficiency,a pair of specific primers for FCV detection were designed based on the conserved region of the reference FCV genome sequences from GenBank with Primer Premier 5.0.The nano-PCR assay was established for FCV detection.The results showed that the nano-PCR assay had good specificity with no cross reaction with other common feline disease viruses.The lowest detectable limit of the assay,which was 10 times higher than that of conventional PCR,was 5.4×10^(-3) pg/μL of the standard template,suggesting that the assay was with good sensitiveness.The sensitivity test was repeated 3 times with good repeatability.In conclusion,the established FCV nano PCR method had the advantages of good specificity,sensitivity and repeatability,which can be used as a powerful technical method for laboratory diagnosis of FCV infection.

关 键 词：猫杯状病毒 呼吸道传染病 纳米PCR 

分 类 号：S852.659.6[农业科学—基础兽医学]