鸽I型副粘病毒的RT-PCR套式PCR检测方法的建立

RT-PCR and Nested PCR for Detection of Pigeon Paramyxovirus Type 1

对 3个中国广东的鸽I型副粘病毒分离株 (P4、P5和P7)基因组RNA以反转录 聚合酶链反应法 (RT_PCR)扩增了F基因 75 %的基因区域 ( 3 4 3～ 1673nt)。其中毒株P4和P7准确扩增出预期大小明亮的DNA片段 ,而P5扩增出片段很淡 ,用该RT_PCR产物作模板以相同引物再作PCR ,结果可以扩增出大小与预期一致的明亮条带。此方法可快速检测鸽I型副粘病毒(PPMV_1)。

RT_PCR was used to amplify 75 percent (from 343nt～1673nt) of the F gene fragment of three Pigeon Paramyxovirus type 1 isolations (P4,P5and P7) from Guangdong province,China.The result indicated the size of the PR_PCR products of P4 and P7 was the same as expected.The RT_PCR products of P5 was very dim.A bright fragment of 1.3kb was abtained by nested PCR Using the same primers when the RT_PCR product of P5 was used for templet.The method could rapidly detect Pigeon Paramyxovirus type 1.