中国动物传染病学报
中國動物傳染病學報
중국동물전염병학보
CHINESE JOURNAL OF VETERINARY PARASITOLOGY
2014年
5期
1-9
,共9页
李仕超%仇旭升%刘开春%周锦萍%刘健%鞠厚斌%谭磊%孙英杰%宋翠萍%刘佩红%丁铲
李仕超%仇旭升%劉開春%週錦萍%劉健%鞠厚斌%譚磊%孫英傑%宋翠萍%劉珮紅%丁鏟
리사초%구욱승%류개춘%주금평%류건%국후빈%담뢰%손영걸%송취평%류패홍%정산
新城疫病毒%鸽源%基因型Ⅵb亚型%F基因%HN基因
新城疫病毒%鴿源%基因型Ⅵb亞型%F基因%HN基因
신성역병독%합원%기인형Ⅵb아형%F기인%HN기인
Newcastle disease virus%pigeon%genotypeⅥb%F gene%HN gene
新城疫是严重危害世界养禽业的一种禽类传染病。近年来，鸽新城疫的流行越来越严重，引起了养殖户和兽医工作者的广泛关注。本研究在4批发病鸽组织病料中分离到了4株鸽源新城疫强毒株和2株鸽源弱毒株。病毒分离株经鸡胚有限稀释法纯化5次后，新鲜尿囊液用于提取病毒RNA。经RT-PCR扩增、测序得到分离株的F基因和HN基因序列。通过对F基因的遗传进化分析发现，4株鸽源强毒株属于ClassⅡ基因Ⅵb亚型，2株鸽源弱毒株与Class II基因II型疫苗株La Sota高度同源。进一步遗传进化分析结果显示，4株鸽源新城疫病毒强毒株与目前国内流行株属于同一遗传分支，这些毒株与欧洲流行株亲缘关系十分相近，而与中国20世纪90年代鸽源分离株遗传距离稍远，因此推测当前鸽源流行强毒株可能源自欧洲。生物信息学分析显示鸽源毒株HN蛋白345~353位的一个线性抗原表位发生了改变，而血清学实验显示La Sota疫苗株和鸽源毒株的交叉血凝抑制存在明显差异，暗示鸽源毒株已经发生了抗原性变异。因此，使用鸡疫苗株La Sota免疫鸽存在免疫失败的风险，需要研发鸽专用新城疫病毒疫苗用于鸽新城疫的防疫工作。
新城疫是嚴重危害世界養禽業的一種禽類傳染病。近年來，鴿新城疫的流行越來越嚴重，引起瞭養殖戶和獸醫工作者的廣汎關註。本研究在4批髮病鴿組織病料中分離到瞭4株鴿源新城疫彊毒株和2株鴿源弱毒株。病毒分離株經鷄胚有限稀釋法純化5次後，新鮮尿囊液用于提取病毒RNA。經RT-PCR擴增、測序得到分離株的F基因和HN基因序列。通過對F基因的遺傳進化分析髮現，4株鴿源彊毒株屬于ClassⅡ基因Ⅵb亞型，2株鴿源弱毒株與Class II基因II型疫苗株La Sota高度同源。進一步遺傳進化分析結果顯示，4株鴿源新城疫病毒彊毒株與目前國內流行株屬于同一遺傳分支，這些毒株與歐洲流行株親緣關繫十分相近，而與中國20世紀90年代鴿源分離株遺傳距離稍遠，因此推測噹前鴿源流行彊毒株可能源自歐洲。生物信息學分析顯示鴿源毒株HN蛋白345~353位的一箇線性抗原錶位髮生瞭改變，而血清學實驗顯示La Sota疫苗株和鴿源毒株的交扠血凝抑製存在明顯差異，暗示鴿源毒株已經髮生瞭抗原性變異。因此，使用鷄疫苗株La Sota免疫鴿存在免疫失敗的風險，需要研髮鴿專用新城疫病毒疫苗用于鴿新城疫的防疫工作。
신성역시엄중위해세계양금업적일충금류전염병。근년래，합신성역적류행월래월엄중，인기료양식호화수의공작자적엄범관주。본연구재4비발병합조직병료중분리도료4주합원신성역강독주화2주합원약독주。병독분리주경계배유한희석법순화5차후，신선뇨낭액용우제취병독RNA。경RT-PCR확증、측서득도분리주적F기인화HN기인서렬。통과대F기인적유전진화분석발현，4주합원강독주속우ClassⅡ기인Ⅵb아형，2주합원약독주여Class II기인II형역묘주La Sota고도동원。진일보유전진화분석결과현시，4주합원신성역병독강독주여목전국내류행주속우동일유전분지，저사독주여구주류행주친연관계십분상근，이여중국20세기90년대합원분리주유전거리초원，인차추측당전합원류행강독주가능원자구주。생물신식학분석현시합원독주HN단백345~353위적일개선성항원표위발생료개변，이혈청학실험현시La Sota역묘주화합원독주적교차혈응억제존재명현차이，암시합원독주이경발생료항원성변이。인차，사용계역묘주La Sota면역합존재면역실패적풍험，수요연발합전용신성역병독역묘용우합신성역적방역공작。
Newcastle disease is a severe infectious disease in birds, threatening poultry industry worldwide. Pigeons are very susceptible to Newcastle disease virus (NDV) infection, which causes high morbidity and mortality. In this study, six NDV strains were isolated from the pigeon tissue samples, four of which were highly virulent and two were low virulent. These pigeon viruses were cloned 5 times in embroynated SPF chicken eggs via limited dilution method. The fresh allantoic fluids were collected for RNA extraction. The resulting RNA samples were converted to cDNAs followed by PCR amplification and sequencing of F and HN genes. Phylogenetic analysis based on F gene sequences showed that four highly virulent strains belonged to ClassⅡⅥb subgenotype and two low virulent strains were similar to La Sota and belonged to ClassⅡgenotypeⅡ. In addition, four highly virulent strains were more closely related to the strain isolated in Italy a few years ago than those strains obtained in China in 1990’s. Therefore, these virulent pigeon strains currently prevailing in China might originate from Europe. Bioinformatics analysis revealed that a linear epitope consisting of amino acids 345-353 on HN protein of these pigeon strains changed as compared with the classic vaccine strain La Sota. In addition, hemagglutination inhibition revealed a noticeable difference between these pigeon strains and La Sota. The results support that chicken vaccine strains do not effectively protect pigeons from NDV infection thus the vaccines designed for pigeon use are much needed.
