Detection of Ilfeetious Laryngotracheitis Virus by Nested PCR

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．