检测感染鸡体内传染性支气管炎病毒反转录套式PCR方法的建立

Establishment of RT Nest PCR for Detection of Infectious Bronchitis Virus in Infected Chicken Tissues

根据先前克隆并测定的鸡传染性支气管炎病毒(IBV)核蛋白(NP)基因序列,设计了2对引物,通过对IBV各毒株(M41,H52,H120,Aust-T以及野毒株等)攻毒鸡的组织中的IBV进行反转录套式PCR检测,结果均为阳性,而对鸡的其他传染病病毒(IBDV,AIV,NDV)进行检测,结果均为阴性。结果表明所建立的反转录套式PCR方法可用于感染鸡体内IBV各血清型毒株的检测。

Two pair of primers to amplify the part of N gene of infectious bronchitis virus(IBV)in the infected chicken were designed and synthesized according to the sequences of prior cloned IBV NP gene . The RT-Nest -PCR were used to detect the IBV in the infected chicken .The positive results were observed from the chicken infected with IBV M41,H52,H120,Aust-T strains and field strain isolated in shanghai , while the negative reaction were showed from the bodies infected with other related avian disease virus( IBDV, AIV, NDV). The results indicated that the established RT-Nest-PCR technique may be used to detect many strains of IBV in the infected bodies.