【作者】 王泽霖； 王宪文； 菅复春； 席瑞珍； 姚惠霞；

【Author】 Wang Zelin Wang Xianwen jian Fuchun Xi Ruizhen Yao Huixia(Avian disease reseach institue,Henan agricultural university Zhengzhou 450002)

【机构】 河南农业大学禽病研究所；

【摘要】 将来自我国5个省的16株IBV地方分离株分别与6株标准血清型毒株(M41Holte、Arka、Conn、Gray、T)在气管环上进行交叉中和试验。结果显示,在我国存在着众多的血清型及其变异株,其中M类有江苏Y、云、郑18、9817、118、Am株;Conn类有118株和Joo株;Gray类有肾-20、E、9801株;Holte类有广30、张IBV、9801株;T类有云、9801株;Ark类有广35、云、郑18;宜类有Am、9801;另有与上述各标准株均无反应的广32株。在16株地方株中.M类呼吸型IBV最多.有6株.占37.5%.仍为当前IBV流行的主要血清型。河南分离的肾变型宜毒株与除Holte株以外的其余标准株均由不同程度的交叉.但与Ark株相关性最大.为25%。将5株标准株和6株地方分离株的0.6kb扩增产物.用三种限制性内切酶Alu I、Taq I和Afa I进行酶切.RFLP分析可将标准株分为如下三种模型①M41模式:Alu I酶切、Taq I酶切均有特异的条带(Afa I酶切均为阳性);②Ark株、G株、C株模式:Alu I酶切阳性而Taq I酶切阴性;③T株模式:Alu I酶切阴性而Taq I酶切阳性。可将各地方株分为两种模式①M41模式有:广32株、J7株、张IBV株、Am株②Ark株、G株、C株模式有138株、宜株;将地方株宜株的0.6kb扩增片段进行测序并与标准株Ark株、G株、C株、Holte株和M41株的相应序列进行分析比较,核苷酸、氮基酸同源性的分析显示.宜毒株与同一酶切类型的Ark-1529-29株同源性最高,核苷酸同源性为93%,氨基酸同源性为91.6%。推测宜株可能是Ark血清型毒株的变异株。血清学分型与基因分型在宜株定位上的一致性,在地方毒株分类上不完全一致性.表明这两种分型方法既有内在联系。又存在着截然不同的差异。更多还原

【Abstract】 Cross neutralization test was done in tracheal ring using sixteen IBV isolates from five province with sixreference serotype strains. It indicates that there are many serotypes and variaants in our country, among which Jiangsu Y, zheng18. 9817,118 and Am belong to M strain pattern; 118 and Joo belong to Conn pattern; Guang30, ZhanglBV , 9801 belong to Holte pattern ; Yun, 9801 belong to T pattern ; Guang35 , Yun , Zheng18 belong to Ark pattern ;Am9801belongs to Yi pattern ;Guang32 is in none of above pattern. Among the sixteen IBV isolates, resperatorypathogenic IBV represented by M41 is the major serotype in the prevailing strains, nephropathogenic Yi strain from Henan province react to all the reference strais except for Holte. It is mostly relevent to Ark(25%). The sequence of the 602bp fragment which contains the part of SI glycoprotein gene of IBV was amplified by polymerase chain reaction (PCR). Five out of six reference strains (M41 .Holte, Arka,Conn,Gray ,T) and six out of eight field isolates(138,Yi,Guang32, Yun,14Hu,Am,J7 ,ZhanglBV) were amplified successfully. We digesed the 602bp fragment of PCR products with three restricton enzyme(RE)AluI , Taq I and Afa I respectively. Six reference strains were grouped into three patterns according to the restriction fragment length polymorphism (RFLP) analysis ①the pattern of M41,digested positively with AluI , Taq I and Afa I ②the pattern of Ark,Gray and Conn digested positively with both Alu I and Afa I but negatively with Taq 1 ③the pattern of T, digested positively with both Taq I and Afa I ,but negatively with Alu I . Similarly, six field strains were grouped into two patterns according to the same restriction fragment length polymorphism analysis :畉he pattern of M41 (Guang32, Am, J7,ZhangIBV),; (2)the pattern of Ark, Gray, Conn (138, Yi). The 602bp fragment of Yi isolate is sequenced and analysed with DNAstar,compared with the relavent sequences of Ark, Gray, Conn which are in the same RFLP pattern and M41 which is in the different RFLP pattern, According to the analysis of homology , Yi isolate share the highest homology with Ark:the neucleutide homology is 93%, the amino acids homology is 91. 6%. It is concluded that Yi isolate may come from the serotype of Ark-1529 -29,or a varriant of it. Serological classification and gene classification is idntical as far as Yi strain is concerned, and partly identical as for field isolates. It indicates that serological differentiiation is internally related but different largely from gene differentiation.更多还原