The use of serotype 1-and serotype 3-specific polymerase chain reaction for the detection of Marek's disease virus in chickens
The use of serotype 1-and serotype 3-specific polymerase chain reaction for the detection of Marek's disease virus in chickens
复制标题
DOI：
10.1080/03079450120054659
复制
发表时间：
2001-06-01
期刊：
影响因子：
2.8
通讯作者：
Jorgensen, PH
中科院分区：
文献类型：
--
作者：
Handberg, KJ;Nielsen, OL;Jorgensen, PH
建立了马立克氏病病毒(MDV)1型和3型特异性聚合酶链式反应(PCR)检测方法。将该方法应用于细胞培养时，培养病毒的灵敏度与培养病毒相当。该方法被应用于实验接种1型或3型MDV的鸡的各种组织样本。分别在接种后56天和84天(P.I.)在羽毛毛囊上皮细胞中检测到血清1型毒株CVI988和RB-1B，而在接种后42天(P.I.)仍可检测到MDV-3型。本研究的目的是建立和评价一种可靠、易操作的监测鸡群马立克氏病发生的方法。我们强调开发一种方法，该方法可以应用于在野外方便地采集的样本类型，例如羽毛尖端和血液样本。此外，还对用1型(CVI988)或3型(HVT)疫苗免疫的4个商品肉用蛋鸡幼鸡或雏鸡群的样本进行了检测。这些鸡群有各种马立克氏病的临床症状。在黄皮细胞、脾、肝、皮肤、羽毛尖和卵巢中检测到MDV-1。在羽毛顶端检测MDV的灵敏度似乎与黄毛细胞共培养一样灵敏，尽管在非白鸡的羽毛顶端提取液中观察到了抑制因子。在大多数提取物中，这种抑制作用可以通过应用牛血清白蛋白预处理来克服。结果表明，该方法是监测家禽中马立克氏病的一种简便方法，羽毛尖端是最方便、最敏感的样本。
A serotype 1- and serotype 3-specific detection of Marek's disease virus (MDV) by polymerase chain reaction (PCR) was developed. The sensitivity of the method when applied to cell culture grown virus was comparable with that of cultivation. The method was applied to various tissue samples from chickens experimentally inoculated with serotype 1 or serotype 3 MDV. The serotype 1 strains CVI988 and RB-1B could be detected in feather follicle epithelium up to 56 and 84 days post-inoculation (p.i.), respectively, while the MDV-3 serotype was detected until 42 days p.i. The purpose of this study was to develop and evaluate a reliable and easy-to-handle method for surveillance of the occurrence of MDV in chicken flocks. We emphasize the development of a method, which can be applied to types of samples conveniently collected in the field, e.g. feather tips and blood samples. In addition, the PCR was applied to samples collected from four commercial table egg layer flocks of young stock or pullets vaccinated with either serotype 1 (CVI988) or serotype 3 (HVT) vaccine. These flocks had various clinical signs of Marek's disease. MDV-1 was detected in buffy-coat cells, spleen, liver, skin, feather tips and ovaries. The detection of MDV in feather tips appeared to be as sensitive as co-cultivation of buffy-coat cells, although an inhibiting factor was observed in extracts from feather tips of non-white chickens. This inhibition could be overcome in most extracts by applying a bovine serum albumen pretreatment. The PCR proved to be a convenient tool for the monitoring of MDV in the poultry population, and feather tips were the most convenient and sensitive samples.