河南农业科学Issue(8)：121-127,140,8.DOI:10.15933/j.cnki.1004-3268.2015.08.027
1株犬细小病毒的分离鉴定及其生物学特性研究
Isolation and Identification of Canine Parvovirus Strain NY130615 and Its Biologic Characterization
摘要
Abstract
The faeces samples were collected from a dog suspected infected by canine parvovirus( CPV) , CPV in the samples was detected by colloidal gold test strip and polymerase chain reaction(PCR),then the samples were innoculated into F81 cells to isolate CPV,propagation dynamics characterization of CPV in F81 cells were determined by immunoperoxidase monolayer assay( IPMA) and PCR,samples that cul-tured with or without serum were titrated by IPMA. VP2 gene of this virus strain was amplified by PCR and sequenced then analyzed by software DNAStar 7 . 0 and MEGA 5 . 0 . Results of colloidal gold test strip test showed CPV Ag negative in the samples,however,PCR detection results showed positive. The sample inoculated into F81 cells showed significant cytopathic effect( CPE) . Serologic test results showed that the culture samples were CPV Ag positive,the isolated virus strain was named CPV-NY130615. CPV Ag in the F81 cells 6 hours post infected( hpi) with CPV-NY130615 strain was successfully detected by IPMA, as well as the culture supernant was CPV nucleotied positive detected by PCR. The fifteenth passages vi-rus titers of two different culture method,synchronization inoculated culture with serum and monolayer in-oculated culture without serum were 1 × 108. 06 TCID50/mL and 1 × 107. 65 TCID50/mL respectively. Nucle-otid sequences showed that the open reading frame of VP2 gene was 1 755 bp encoding 584 aa. Phyloge-netic analysis showed that CPV-NY130615 strain belonged to the genotype CPV-2a.关键词
犬细小病毒/分离/免疫过氧化物酶单层细胞染色法/生物学特性Key words
canine parvovirus/isolation/immunoperoxidase monolayer assay/biological characteristics分类
农业科技引用本文复制引用
卜宾,李生涛,毛倩倩,唐青海,唐存多,焦铸锦,姚伦广,阚云超..1株犬细小病毒的分离鉴定及其生物学特性研究[J].河南农业科学,2015,(8):121-127,140,8.基金项目
国家自然基金青年基金项目(31101837) （31101837）
河南省重点科技攻关项目(142102110101) （142102110101）
南阳师范学院引进人才专项(70640) （70640）
南阳师范学院研究生创新基金项目(20141101,2014103) （20141101,2014103）
