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一株鸽源Ⅰ型副黏病毒的分离鉴定和F基因序列分析

Isolation and identification of an isolate of pigeon Paramyxovirus type 1 and sequence analysis of F gene

  • 摘要: 为了研究山西地区鸽源Ⅰ型副黏病毒(PPMV-1)的遗传进化规律,试验通过采集疑似病鸽临床病料,经SPF鸡胚传代分离病毒,采用血凝试验和RT-PCR对分离株进行鉴定,扩增分离株F基因部分片段进行同源性分析,并通过病毒毒力测定试验和动物回归试验评价分离株的毒力和致病性。结果表明:从发病鸽群中分离鉴定1株PPMV-1,命名为PPMV-1-SX-011,分离株F基因裂解位点氨基酸序列为112RRQKRF117,具有新城疫病毒(NDV)强毒株的典型分子特征;分离株PPMV-1-SX-011与国内流行株Pi/CH/LN/2017、Pi/SH/CH/0168/2013、Pigeon/China/BJ2013、pigeon/China/SDS/2011在遗传进化树上处于同一分支,其中与Pi/SH/CH/0168/2013、pigeon/China/SDS/2011的核苷酸序列同源性高达98.1%和97.7%,属于ClassⅡ基因Ⅵb型;但是与经典疫苗株La Sota、B1、Clone30和Mukteswar株的同源性仅为82.7%~84.5%。分离株PPMV-1-SX-011最小致死量致死鸡胚平均死亡时间(MDT)、1日龄雏鸡脑内接种致病指数(ICPI)、6周龄SPF鸡静脉接种致病指数(IVPI)值分别为105.6 h、0.3和0。分离株能引起试验鸡和试验鸽体内的免疫应答反应;对试验鸽的致病力强,而对试验鸡无致病力,表明分离株对鸽有特殊亲嗜性。说明本研究分离得到一株鸽源PPMV-1,其具有NDV强毒株裂解位点,对鸡的致病力却与传统NDV存在明显差异,提示山西地区存在对鸽具有强致病性的NDV弱毒株的基因变异株。

     

    Abstract: In order to study the genetic evolution of the pigeon Paramyxovirus type 1(PPMV-1) in Shanxi Province, by collecting clinical disease materials of suspected sick pigeons, the virus was isolated through SPF chicken embryo passage, and the isolated strains were identified by hemagglutination test and RT-PCR. Some fragments of F gene of the isolates were amplified for homology analysis. The virulence and pathogenicity of the isolates were evaluated through virus virulence test and animal regression test. The results showed that one strain of PPMV-1 was isolated and identified from the diseased pigeon group, named PPMV-1-SX-011; the amino acid sequence of the cleavage site of the F gene of the isolate was112RRQKRF117, which had the typical molecular characteristics of the virulent NDV strain; the isolate PPMV-1-SX-011 and the domestic popular strains Pi/CH/LN/2017, Pi/SH/CH/0168/2013, Pigeon/China/BJ2013, pigeon/China/SDS/2011 belonged to the same branch on the genetic evolution tree, and the nucleotide sequence homology of PPMV-1-SX-011 with Pi/SH/CH/0168/2013, pigeon/China/SDS/2011 was 98.1% and 97.7%, belonging to ClassⅡ genotype Ⅵb; however, its the homology with the classic vaccine strains La Sota, B1, Clone30 and Mukteswar strains was only 82.7%-84.5%. The values of the average death time(MDT) of embryos being resulted in by the minimum lethal dose, the pathogenicity index of intracerebral vaccination(ICPI) of one-day-old chicks, and the pathogenicity index of intravenous vaccination(IVPI) of six-week-old SPF chickens of the isolate PPMV-1-SX-011, were 105.6 h, 0.3 and 0, respectively. The isolate could cause an immune response in chicken and pigeon; it was highly pathogenic to pigeon, but not to chicken, indicating that the isolate had a special affinity for pigeon. The results suggested that in this study, a pigeon-derived PPMV-1 was isolated, which had a cleavage site for the virulent strain of NDV, but its virulence to chicken was different from that of traditional NDV, which indicated that there were genetic variants of NDV attenuated strain with strong pathogenictity to pigeons in Shanxi Province.

     

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