RT—PCR对新城疫病毒（NDV）分离株的鉴定及其临床应用

采用新城疫病毒的通用引物PA PB、强毒引物PA PC、弱毒引物PA PD对11株贵州新城疫分离毒株与4株新城疫参考毒株进行了RT-PCR扩增.结果15株新城疫毒株均被PA PB引物扩增出359 bp的条带,F48E8、ND98、Fw、H2、P3、BY、L2、P1,P2被PA PC引物扩增出254 bp的条带,而PA PD引物未扩增出DNA条带;ND89、Lasota被PA PD引物扩增出254 bp的条带.而PA PC引物未扩增出DNA条带.采用3对引物对自然发病斗鸡脑、脾、咽喉试子、泄殖腔试子进行RT-PCR检测,确诊为斗鸡新城疫强毒感染,且与传统病毒分离与毒力鉴定的结果相一致.

Identification of Newcastle disease virus(NDV)isolates by RT-PCR and its clinical application

RT-PCR assays have been conducted to differentiate NDV isolates by using 3 pairs of primers,which were designed according to the gene sequence at the cleavage site of F protein.PA PB primers directed at all NDV strains;PA PC,PA PD primers were specific to virulent and non-virulent strains respectively.The results showed a fragment of 359 bp was amplified by PA PB primers from 15 strains of NDV isolates.The RT-PCR by using PA PC primers resulted in a fragment of 254 bp from F48E8,ND98,Fw,H2,P3,BY,L2,P1,P2 strain and no band was found by using PA PD primers.Meanwhile ND89,Lasota strains produced a fragment of 254 bp by using PA PD primers and manifested negative reaction by using PA PC primers.The brain,spleen,pharyngeal and cloacal swabs were sounpled from naturally infected cockfights and detected with RT-PCR assays by using the 3 pairs of primers.The sick cockfights were definitely diagnosed as virulent NDV infection.It is proved to be in accordance with the traditional methods of virus isolation and virulence identification.