Virology Unit, Institute for Microbiology and Parasitology, Veterinary Faculty, University of Ljubljana, Gerbiceva 60, SI-1115 Ljubljana, Slovenia.
J Virol Methods. 2010 Jun;166(1-2):28-36. doi: 10.1016/j.jviromet.2010.02.012. Epub 2010 Feb 17.
Newcastle disease virus (NDV), also designated avian paramyxovirus type 1 (APMV-1), is a serious pathogen of poultry, causing highly contagious Newcastle disease (ND), with high morbidity or mortality, depending on the strain. Accordingly, rapid and reliable detection of APMV-1 and differentiation between vaccine and virulent strains is of crucial importance for ND diagnosis and plays an important role in effective control of the disease. In this study, two real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assays using minor groove-binding (MGB) probes were developed for broad range detection and simultaneous pathotyping of APMV-1. The two assays were evaluated for their ability to detect in allantoic fluids viral RNA of all known APMV-1 lineages. Additionally, the applicability of the developed assays was assessed by the detection and pathotype prediction of APMV-1 in swabs and organs. The assays demonstrated high analytical specificity, sensitivity and good reproducibility, with coefficients of variation ranging from 0.2% to 3.9% and from 0.6% to 7.2% for intra-assay and inter-assay variability, respectively. The results indicated the suitability of both assays as a complementary method for rapid screening and typing of APMV-1.
新城疫病毒（NDV），也称为禽副黏病毒 1 型（APMV-1），是家禽的一种严重病原体，可引起高度传染性的新城疫（ND），其发病率或死亡率取决于毒株。因此，快速可靠地检测 APMV-1 并区分疫苗株和强毒株对于 ND 的诊断至关重要，对有效控制疾病也具有重要作用。在本研究中，开发了两种基于小沟结合（MGB）探针的实时逆转录聚合酶链反应（real-time RT-PCR）检测方法，用于广泛检测和同时对 APMV-1 进行定型。评估了这两种检测方法对所有已知 APMV-1 谱系的鸡胚尿囊液病毒 RNA 的检测能力。此外，还通过对拭子和器官中 APMV-1 的检测和预测，评估了开发检测方法的适用性。结果表明，这两种检测方法均适合作为 APMV-1 的快速筛选和定型的补充方法。
