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《Journal of Northwest A & F University(Natural Science Edition)》 2011-01
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Establishment and application of identification methods of Nephropathogenic IBV strain and respiratory-type vaccine strain

XU Xue1,GAO Dong-sheng1,WANG Chuan-qing1,CHEN Lu1,YANG Xia1,HUANGFU He-ping1,2,LIU Hui-min1,ZHENG Lu-ping1,YANG Meng1,ZHANG Jin-lai1,LI Kun-ming1,ZHANG Jiu-zhou1 (1 Institute of Poultry Diseases,Henan Agricultural University,Zhengzhou,Henan 450002,China;2 Zhengzhou College of Animal Husbandry Engineering,Zhengzhou,Henan 450011,China)  
【Objective】Based on the S1 gene of IBV,the reverse transcription-multiple PCR methods were established to differentiate the epidemic Nephropathogenic strains and respiratory-type vaccine strains of IBV.【Method】 According to the conservative nucleocapsid gene sequences of avian infectious bronchitis virus(IBV),a pair of specific primers was designed and synthesized.And on this basis the molecular biological detection methods of IBV were established to assay the IBV isolates.At the same time,by analyzing the complete sequences of the IBV S1 gene published on GenBank,the relatively conservative differential sequences of respiratory-type vaccine strain and local nephropathogenic epidemic strains were found out.Two pairs of primers were designed and synthesized and the reverse transcription-multiple PCR methods were established to type for the two serotypes of IBV.Simultaneously,the method whose sensitivity and reproducibility had been tested was applied to clinical detection.【Result】 The typing method was established successfully.The genome RNA diluted to 10-6 could still be amplified effectively using this method.The results showed that:the methods could distinguish respiratory-type strains from the local nephropathogenic strains isolated in recent years better;the nephropathogenic strains(YI strain and the Australian T strain) isolated in the earlier period were not amplified effectively;in recent years,the strains that could infect chickens and cause high mortality mainly were nephropathogenic IBV.【Conclusion】 The typing method distinguishing local nephropathogenic epidemic strains from respiratory-type vaccine strains of IBV was established.The method has characteristics of strong specificity,high sensitivity and good repetition.And the method can distinguish completely the local nephropathogenic IBV from the respiratory-type IBV vaccine strains,and supply reference for detection and vaccine options of IB.
【Fund】: 中德合作项目“Monitoring the prevalence and significance of Gallibacteriumanatis in Chinese poultry flocks”(43006167)
【CateGory Index】: S858.316.3
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