Jackwood D J, Nielsen C K
Ohio State University, Wooster 44691, USA.
Avian Dis. 1997 Jan-Mar;41(1):137-43.
Bursa samples from the United States, Mexico, and Puerto Rico were tested for the presence of infectious bursal disease virus (IBDV) using the reverse transcriptase/polymerase chain reaction-restriction endonuclease (RT/PCR-RE) assay. This assay amplifies a 394-bp fragment of the IBDV VP2 gene. A total of 151 samples were tested. Each was from a different physical location or farm. Forty-eight of the samples were determined to contain IBDV using RT/PCR. The RE profiles on 44 of these positive samples were determined using the enzymes BstNI or EcoRII, StyI, DraI, SacI, Sau3AI or MboI, and TaqI. A majority of the samples (34) had RE profiles typical of variant IBDV strains. One of six samples from Mexico was positive for IBDV. This virus had an RE profile typical of variant strains of IBDV. Three of seven samples from Puerto Rico had RE profiles characteristic of variant viruses. Two samples from the United States had RE profiles characteristic of classic vaccine IBDV strains and nine samples had new RE profiles. Five of these new profiles were BstNI- and StyI-negative, indicating that these viruses may be antigenically related to variant types. Although the new RE pattern observed in the other four samples was BstNI- and StyI-positive, it was not typical of classic vaccine IBDV strains. One flock from the United States had a mixture of two RE profiles, a typical variant type profile and an unknown variant RE profile. Two-thirds of the positive samples from flocks where the age of the birds was reported were observed between 21 and 28 days of age. The results of these studies demonstrate that the RT/PCR-RE assay can be used to diagnose IBDV in chickens and that IBDV strains exist in commercially reared chickens that have RE patterns different than known IBDV strains. The molecular differences observed using the RT/PCR-RE test were in a region of the VP2 gene, which is known to code for important neutralizing epitopes and to be highly variable among IBDV strains. Although the results demonstrate RE patterns different than those observed in known classic and variant IBDV strains, the influence of these molecular differences on biological properties of the viruses requires further investigation.
使用逆转录酶/聚合酶链反应-限制性内切酶（RT/PCR-RE）分析法对来自美国、墨西哥和波多黎各的法氏囊样本进行传染性法氏囊病病毒（IBDV）检测。该分析法可扩增IBDV VP2基因的一个394碱基对片段。共检测了151个样本。每个样本都来自不同的地理位置或养殖场。使用RT/PCR法确定其中48个样本含有IBDV。使用BstNI或EcoRII、StyI、DraI、SacI、Sau3AI或MboI以及TaqI酶确定了其中44个阳性样本的RE图谱。大多数样本（34个）具有变异IBDV毒株典型的RE图谱。来自墨西哥的六个样本中有一个IBDV呈阳性。该病毒具有IBDV变异毒株典型的RE图谱。来自波多黎各的七个样本中有三个具有变异病毒特征性的RE图谱。来自美国的两个样本具有经典疫苗IBDV毒株特征性的RE图谱，九个样本具有新的RE图谱。其中五个新图谱对BstNI和StyI呈阴性，表明这些病毒可能在抗原性上与变异类型相关。尽管在其他四个样本中观察到的新RE模式对BstNI和StyI呈阳性，但它并非经典疫苗IBDV毒株的典型模式。来自美国的一个鸡群有两种RE图谱的混合，一种是典型的变异型图谱，另一种是未知的变异RE图谱。在报告了鸡龄的鸡群中，三分之二的阳性样本是在21至28日龄之间观察到的。这些研究结果表明，RT/PCR-RE分析法可用于诊断鸡的IBDV，并且在商业饲养的鸡中存在具有与已知IBDV毒株不同RE模式的IBDV毒株。使用RT/PCR-RE检测观察到的分子差异存在于VP2基因的一个区域，该区域已知编码重要的中和表位，并且在IBDV毒株之间高度可变。尽管结果表明RE模式与已知的经典和变异IBDV毒株中观察到的不同，但这些分子差异对病毒生物学特性的影响需要进一步研究。
