Keam L, York J J, Sheppard M, Fahey K J
Commonwealth Scientific and Industrial Research Organisation, Division of Animal Health, Parkville, Victoria, Australia.
Avian Dis. 1991 Apr-Jun;35(2):257-62.
A DNA hybridization assay using a non-radioactive probe has been developed for the detection of infectious laryngotracheitis virus (ILTV) DNA. A 1.4-kilobase pair BamHI fragment of ILTV genomic DNA was cloned and then labeled by one of two methods; nick translation using 32P-dATP or non-radioactive labeling using a commercially available DNA labeling and detection kit. The non-radioactive DNA labeling method proved to be as sensitive as the radioactive method. Using the non-radioactive probe, ILTV DNA was readily detected in tracheal samples from acutely infected chickens and also from convalescent chickens at a time when viral antigen could no longer be detected by the enzyme-linked immunosorbent assay or the virus could no longer be reisolated. This technique provides a safe and effective means of identifying field outbreaks of ILTV and also may detect latent ILTV infections relatively quickly and inexpensively.
已开发出一种使用非放射性探针的DNA杂交检测方法，用于检测传染性喉气管炎病毒（ILTV）DNA。克隆了ILTV基因组DNA的一个1.4千碱基对的BamHI片段，然后通过两种方法之一进行标记：使用32P-dATP进行缺口平移或使用市售的DNA标记和检测试剂盒进行非放射性标记。结果证明，非放射性DNA标记方法与放射性方法一样灵敏。使用非放射性探针，在急性感染鸡的气管样本中很容易检测到ILTV DNA，在恢复期鸡的气管样本中也能检测到，此时酶联免疫吸附试验已无法检测到病毒抗原，也无法再分离出病毒。这项技术提供了一种安全有效的方法来识别ILTV的田间疫情，还可能相对快速且廉价地检测出潜伏的ILTV感染。
