马源禽Ⅰ型副粘病毒的分离与鉴定

Isolation and identification of avian paramyxovirus serotypeⅠ from equine

【目的】了解广西马源Ⅰ型副粘病毒的生物特性,为今后有效防控副粘病毒在不同宿主间传播提供科学依据。【方法】用SPF鸡胚对广西百色疑似病马组织悬浮液进行病原分离培养后,经血清学试验和致病性试验鉴定,再用RT-PCR对分离株F基因进行扩增,并对扩增片段进行测序分析。【结果】分离获得的病毒能使鸡红细胞发生凝集,且这种凝集可被鸡新城疫(ND)标准阳性血清所抑制,HA和HI效价分别为27和210。分离株毒力试验结果显示,鸡胚平均死亡时间(MDT)为72 h,雏鸡脑内接种致死指数(ICPI)为1.48,属中发型毒株。分离株与禽I型副粘病毒基因Ⅶ型毒株NDV04-21、TW-96p的核苷酸同源性分别为95.9%和95.1%;基于F基因的遗传分型进化树显示,分离株与NDV04-21、TW-96p处于同一进化树分支,属于禽I型副粘病毒基因Ⅶ型;分离株F蛋白112～117裂解位点氨基酸序列与标准强毒株F48E9、HER33的一致,在第112～117位氨基酸序列为3'-R-R-Q-R-R-F-5'。【结论】广西马群已有禽I型副粘病毒存在,再次证实广西地区禽I型副粘病毒宿主范围在不断扩大。

[Objective]This study was aimed to understand the biological characteristics of horse origin avian paramyxovirus serotype Ⅰ and provide references for preventing and controlling the paramyxovirus interspecies transmission.[Method] In Baise of Guangxi,causative agents were isolated from suspicious samples of horse using chicken embryos inoculation and cultured.After serological experiment and pathogenicity identification,F gene of isolates was amplified using RT-PCR and sequenced.[Result]The isolated strain could agglutinate chicken erythrocytes and was inhibited by Newcastle disease virus antiserum.Its titer of HA and HI was 27 and 210,respectively.The MDT and ICPI of chicken embryos was 72 h and 1.48,respectively.The isolated strains,which was moderate susceptible,belonged to genotype Ⅶ,sharing 95.9％ and 95.1％ nucleotide sequence homologies with NDV04-21 and TW-96p,respectively.According to the phylogenetic tree of F gene,the isolates,NDV04-21 and TW-96p belonged to the same branch,which were APMV-Ⅰ genotype Ⅶ.Amino acid sequence of the isolate fusion （F） protein cleavage site 112-117 was 3＇-R-R-Q-K-R-F-5＇,which was in accordance with standard virulent strain HER33 and F48E9.[Conclusion]The results indicated that APMV-Ⅰ infected horses.It was reconfirmed that hosts of APMW-1 were developing in Guangxi area.