选择鸡传染性喉气管炎病毒保守TK基因的蛋白编码区域，设计并合成了一对外引物和一对内引物，建立并 优化了检测鸡传染性喉气管炎病毒DNA的套式PCR法。通过检测IⅡv感染的鸡胚绒毛尿囊膜、实验室病料和临 床病料，结果表明，套式PCR法能检测出ILTV感染后的非免疫鸡胚和SPF鸡胚绒毛尿囊膜研磨液中的被稀释了 105倍的病毒(约1 的Ⅱ v DNA)，攻毒后第10天还能从非免疫鸡和SPF鸡气管拭子中检出Imv，第1O天非免疫 鸡气管拭子中Ⅱ爪，的最大检出率为7／10，第10天SPF鸡气管拭子中IⅢ 的最大检出率为8／10。对非免疫鸡和 SFF鸡的气管拭子中IⅡv最佳检出时间均在攻毒后第5天。对临床样品中的IⅡv的最大检出率为7／7。经过核 酸杂交验证，套式pcR法具有很高的特异性和敏感性．为从分子水平探讨Ⅱ v的发病机理、临床早期快速诊断提 供了新的研究手段

Two pairs of oligonucleotides flanking 33aymidine Kinase(TK)seonent of infectious laryngotracheifs virus(Ⅲ )were chosen as primers for polymerase chaln reaction(nested PCRj． e method of nested PCR W~LS．used to detect of virus DNA in ILTV infected diferent chorlollantoic membrane，specimens from laboratoO, and 7 clinical specimens from various species， m ILTV gene TK seglllent p1 d DNA as positive contro1． The vlms DNA was extracted by acid guanidinium thlocyanate-phenol—chloroform slngle-step method ．Th e re— suhs sho,a-ed that allⅡ V infected diferent chorloUantoic memb rane were positive．The highest detection rate ofⅡ Ⅳ from tracheal swabs of Iqon—hnmmtized chicken was 7／10 on the tenth day P．i．．and that from tracheal swal3~of SPF chickens was 8／10 on the tenth day P．i．．The best time for唧detection in tracheal swabs fmm both rloii—inmmnized chickens and SPF chickens wasthefifth day P．i． detectionfrom clinical 蚋m— ples by nested PCR W~LS．at a rate of7／7．To~,erify the nucleotide hybridization test th TKc probe ．it suggests that nested PCR is sensitive，specific，rapid mad simple，and can be印 ed for study of pathogenesis on moiec— ular level and for early clinical diagnosis．