PCR制备地高辛标记的探针检测禽流感病毒核酸

用聚合酶链反应（PCR）技术，制备了广东禽流感无致病力分离株A/goose/China/24/96(H7N3）核蛋白基因片段（NPc）的地高辛标记的cDNA探针。建立并优化了检测禽流感病毒核酸的探针杂交法，探针杂交法能鉴别出非免疫鸡胚和SPF鸡胚尿囊液中的病毒，攻毒后第3天的SPF和非免疫鸡泄殖腔拭子中AIV的最大检出率为1/10，对临床样品中的AIV的最大检出率为1/7，而直接HA和HI法及AGP试验检不出临床样品的AIV。该探针具有较好的特异性和敏感性，为从分子水平探讨AIV的发病机理、临床早期快速诊断提供了新的研究手段。

Detection of nucleic acid of AIV by a dig-labelled probe prepared by polymerase chain reaction (PCR)

A cDNA probe of the partial genome segment of avian influenza virus (AIV) was labelled with DIG by the technique of PCR. It was used for detection of virus RNA in AIV infected chicken embryo allantoic fluids clinical specimens from various avian. The positive rate of cloacal swab from non immunized chickens and SPF chickens experimentioly infected with AIV were 10%(1/10) by dot hybridization on the third day p.i.. It was 14.3%(1/7) for clinical samples. However, direct HA, HI AGP test could not detect AIV in these clinical smples. This showed that the methods of dot hybridization established were more sensitive, rapid and accurate than conventional serologic test. It can be applied for study on the pathogenesis in molecular level.