Avian Disease Division, Animal and Plant Quarantine Agency, GimCheon, Republic of Korea.
Avian Disease Division, Animal and Plant Quarantine Agency, GimCheon, Republic of Korea.
Poult Sci. 2021 Apr;100(4):100986. doi: 10.1016/j.psj.2021.01.009. Epub 2021 Jan 16.
Infectious laryngotracheitis (ILT), fowlpox (FP), and reticuloendotheliosis are important poultry diseases caused by gallid herpesvirus 1 (ILTV), fowlpox virus (FWPV), and reticuloendotheliosis virus (REV), respectively. Coinfections with ILTV and FWPV occur naturally in chickens, and FP in its more virulent wet form is characterized by diphtheritic lesions and easily confused with ILT. Moreover, the insertion of only partial REV-LTR or a nearly full-length REV into the FWPV genome, located between the ORF 201 and ORF 203, has increased recently in wild-type field FWPV isolates. Therefore, it is critical to detect ILTV, FWPV, REV-integrated FWPV, and REV early and accurately. In this study, we successfully developed a multiplex PCR assay for the simultaneous detection of ILTV, FWPV, REV-integrated FWPV, and REV, and the detection limits was 1 × 5 copies/tube. When used to test clinical samples, the results of the multiplex PCR were in 100% agreement with singleplex PCRs and sequencing. This new multiplex PCR is a simple, rapid, sensitive, specific, and cost-effective method for detection of 4 viruses in clinical specimens.
传染性喉气管炎（ILT）、禽痘（FP）和网状内皮组织增殖症是由禽疱疹病毒 1（ILTV）、禽痘病毒（FWPV）和网状内皮组织增殖症病毒（REV）分别引起的重要家禽疾病。ILTV 和 FWPV 的混合感染在鸡中自然发生，湿性形式的 FP 以白喉样病变为特征，容易与 ILT 混淆。此外，REV-LTR 的部分插入或几乎全长的 REV 插入到位于 ORF201 和 ORF203 之间的 FWPV 基因组中，在野生型田间 FWPV 分离株中最近有所增加。因此，早期、准确地检测 ILTV、FWPV、整合了 REV 的 FWPV 和 REV 至关重要。在本研究中，我们成功开发了一种用于同时检测 ILTV、FWPV、整合了 REV 的 FWPV 和 REV 的多重 PCR 检测方法，检测限为 1×5 拷贝/管。当用于测试临床样本时，多重 PCR 的结果与单重 PCR 和测序完全一致。这种新的多重 PCR 是一种用于检测临床标本中 4 种病毒的简单、快速、敏感、特异和具有成本效益的方法。
