Pedersen J C, Senne D A, Panigrahy B, Reynolds D L
National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Ames, IA 50010, USA.
Avian Dis. 2001 Jul-Sep;45(3):581-92.
Conventional nested and TaqMan reverse transcription-polymerase chain reaction (RT-PCR) assays for the detection of avian pneumovirus (APV) were evaluated and compared with virus isolation (VI) for sensitivity and specificity. Respiratory tissues and tracheal swabs were collected from experimentally inoculated turkeys between 1 and 21 days postinoculation (DPI) and tested by all detection methods. APV was detected by both RT-PCR procedures as early as 1 DPI and as late as 17 DPI, whereas virus was isolated only between 3 and 7 DPI. Pooled tracheal swab supernatant and dry swabs were excellent specimens for the detection of APV between 3 and 8 DPI. Turbinate and sinus specimens were the most productive samples over the entire collection period. Both RT-PCR assays were rapid and more sensitive than VI for the detection of APV in tissue and swab specimens from infected turkeys. RT-PCR allows for the rapid detection of APV from a variety of respiratory tissues as well as from dry swabs and tracheal swab supernatants. Antibody to APV was detected in 50% of the sampled APV-inoculated birds at 8 and 9 DPI by enzyme-linked immunosorbent assay (ELISA). Early seroconversion (8-10 DPI) allows antibody detection to be used as a screening tool for APV. Rapid and sensitive detection methods are needed for APV, a highly contagious disease affecting U.S. poultry.
对用于检测禽肺病毒(APV)的传统巢式和TaqMan逆转录聚合酶链反应(RT-PCR)检测方法进行了评估,并与病毒分离(VI)法在敏感性和特异性方面进行了比较。在接种后1至21天(DPI),从经实验接种的火鸡中采集呼吸道组织和气管拭子,并采用所有检测方法进行检测。两种RT-PCR方法最早在1 DPI就能检测到APV,最晚在17 DPI能检测到,而病毒仅在3至7 DPI能分离出来。合并的气管拭子上清液和干拭子是在3至8 DPI检测APV的优良标本。鼻甲和鼻窦标本在整个采集期内是最易检测到病毒的样本。两种RT-PCR检测方法在检测感染火鸡的组织和拭子标本中的APV时,都比病毒分离法快速且更灵敏。RT-PCR能够从多种呼吸道组织以及干拭子和气管拭子上清液中快速检测出APV。在8和9 DPI时,通过酶联免疫吸附测定(ELISA)在50%的采样APV接种鸡中检测到了APV抗体。早期血清转化(8 - 10 DPI)使得抗体检测能够用作APV的一种筛查工具。APV是一种影响美国家禽的高度传染性疾病,因此需要快速且灵敏的检测方法。