通过简化的聚合酶链反应程序对传染性喉气管炎与其他禽类呼吸道疾病进行鉴别诊断。

Differential diagnosis of infectious laryngotracheitis from other avian respiratory diseases by a simplified PCR procedure.

作者信息

Scholz E, Porter R E, Guo P

机构信息

Department of Veterinary Pathobiology, Purdue Cancer Center, Purdue University, West Lafayette, IN 47907.

出版信息

J Virol Methods. 1994 Dec;50(1-3):313-21. doi: 10.1016/0166-0934(94)90186-4.

DOI:10.1016/0166-0934(94)90186-4

PMID:7714054

Abstract

A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of avian infectious laryngotracheitis virus (ILTV) in chicken trachea, chorio-allantoic membrane (CAM), infected hepatoma cells and infectious cell culture supernatant. Samples were prepared by dilution in distilled water. After boiling and low speed centrifugation, samples were used for PCR analysis with two primers without special labeling. The PCR analysis for ILT virus could be completed in less than 8 h. Standard agarose gel electrophoretic analysis of the PCR products revealed a prominent band of 300 base-pairs in samples from ILTV-infected specimens, but not from specimens containing Newcastle disease virus, infectious bronchitis virus, avian adenovirus, fowlpox virus, Pachecoz or Marek's disease virus. One single ILTV infected cell or 10 plaque forming units of ILTV could be detected with this procedure. The procedure can be used for the identification of ILTV and the differentiation of ILTV from other avian respiratory tract infectants.

摘要

开发了一种基于简单聚合酶链反应（PCR）的方法，用于检测鸡气管、绒毛尿囊膜（CAM）、受感染的肝癌细胞和传染性细胞培养上清液中的禽传染性喉气管炎病毒（ILTV）。样品通过用蒸馏水稀释制备。煮沸并低速离心后，样品用于使用两种无特殊标记的引物进行PCR分析。ILT病毒的PCR分析可在不到8小时内完成。PCR产物的标准琼脂糖凝胶电泳分析显示，来自ILTV感染标本的样品中有一条突出的300个碱基对的条带，但来自含有新城疫病毒、传染性支气管炎病毒、禽腺病毒、禽痘病毒、帕切科氏病或马立克氏病病毒的标本中没有。用该方法可以检测到一个单个的ILTV感染细胞或10个ILTV蚀斑形成单位。该方法可用于ILTV的鉴定以及ILTV与其他禽呼吸道感染因子的区分。