禽腺病毒-Ⅰ群PCR-RFLP分型方法的建立  

Establishment of PCR-RFLP diagnosis method for differentiate group Ⅰ fowl adenovirus

作　　者：江之瑶 王守春[1] 栾庆东 尹燕博[1] 王建琳[1] JIANG Zhi-yao;WANG Shou-chun;LUAN Qing-dong;YIN Yan-bo;WANG Jian-lin(Collage of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109

出　　处：《中国兽医杂志》2019年第11期22-24,27,共4页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划项目(2016YFD0500800)。

摘　　要：为快速确定临床禽腺病毒-Ⅰ群(FAdVs)的血清型,本研究根据FAdVs的Hexon基因设计引物,对12个血清型FAdVs标准毒株进行PCR扩增,并根据扩增目的片段进行酶切位点分析和相应酶切反应研究,建立聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法。结果显示,利用该引物可同时扩增12个血清型FAdVs标准毒株,扩增片段大小为894 bp;扩增片段酶切位点分析显示,Eco130Ⅰ、Pf123Ⅱ、MluⅠ、PsyⅠ和BgⅡ为主要酶切位点,这5种酶切反应结果显示12个血清型FAdVs标准毒株可以得到不同的酶切图谱,且能区分12种血清型FAdVs;应用该酶切方法对本实验室保存的2株已知血清型的分离株进行分析,其结果与已确定的血清型一致。表明本试验建立的PCR-RFLP方法简便、特异性和区分度好,可用于12个血清型FAdVs的分型。In order to differentiate group Ⅰ fowl adenovirus quickly, the universal primers were designed based on the highly conserved Hexon gene of fowl adenosis standard strain.The cleavage sites were designed on the amplification sequence, and the amplified fragments were cut into fragments with different size by restriction endonucleases to establish the method of polymerase chain reaction-restriction fragment length polymorphism. The results showed that the fragments of Hexon for 12 serotype of group Ⅰ fowl adenovirus were successfully amplified, and Eco130 Ⅰ、Pf123Ⅱ、Mlu Ⅰ 、Psy Ⅰ and BgⅡwere the main sites of restriction endonucleases. After being reacted with the five restriction endonucleases, the products of amplified were cut into fragments with different size for different serotype of virus. Two isolated strains of FAdVs-4 and FAdVs-11 were analyzed by the method established in the study, and the results were consistent with that of gene sequencing. The results suggest that polymerase chain reaction-restriction fragment length polymorphism is established in the study with the characteristic of simple, accurate and high differentiation for distinguish the serotype of group Ⅰ fowl adenovirus.

关 键 词：禽腺病毒-Ⅰ群 PCR-RFLP 限制性内切酶 分型 HEXON 

分 类 号：R511.8[医药卫生—内科学]