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首页> 外文期刊>Avian Diseases >Development of a Polymerase Chain Reaction to Differentiate Avian Leukosis Virus (ALV) Subgroups: Detection of an ALV Contaminant in Commercial Marek's Disease Vaccines
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Development of a Polymerase Chain Reaction to Differentiate Avian Leukosis Virus (ALV) Subgroups: Detection of an ALV Contaminant in Commercial Marek's Disease Vaccines

机译：差异化禽白血病病毒（ALV）亚组的聚合酶链反应的发展：商业马立克氏病疫苗中ALV污染物的检测。

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Avian leukosis viruses (ALVs) are common in many poultry flocks and can be detected using an enzyme-linked immunosorbent assay or any other test designed to identify p27, the group-specific antigen located in gag. However, endogenous retroviruses expressing p27 are often present and can be confused with exogenous ALVs. A more specific and informative assay involves targeting the variable envelope glycoprotein gene (gp85) that is the basis for dividing ALVs into their different subgroups. We designed polymerase chain reaction (PCR) primers that would specifically detect and amplify viruses from each of the six ALV subgroups: A, B, C, D, E, and J. Subgroup B and D envelopes are related, and our B-specific primers also amplified subgroup D viruses. We also designed a set of common primers to amplify any ALV subgroup virus. To demonstrate the usefulness of these primers, we obtained from the Center for Veterinary Biologics in Iowa culture supernatant from chicken embryo fibroblasts infected with an ALV that was found to be a contaminant in two commercial Marek's disease vaccines. Using our PCR primers, we demonstrate that the contaminant was a subgroup A ALV. We cloned and sequenced a portion of the envelope gene and confirmed that the ALV was a subgroup A virus. Unlike typical subgroup A viruses, the contaminant ALV grew very slowly in cell culture. We also cloned and sequenced a portion of the long terminal repeat (LTR) from the contaminant virus. The LTR was found to be similar to those LTRs found in endogenous ALVs (subgroup E) and very dissimilar to LTRs normally found in subgroup A viruses. The E-like LTR probably explains why the contaminant grew so poorly in cell culture.
机译：禽白血病病毒（ALV）在许多家禽群中都很常见，可以使用酶联免疫吸附测定法或任何其他旨在鉴定p27（位于gag中的组特异性抗原）的测试方法进行检测。但是，经常存在表达p27的内源性逆转录病毒，并且可以与外源ALV混淆。一种更具体且信息更丰富的测定法涉及靶向可变包膜糖蛋白基因（gp85），这是将ALV分为不同亚组的基础。我们设计了聚合酶链反应（PCR）引物，可特异性检测和扩增六个ALV子组（A，B，C，D，E和J）中的每一个的病毒。B和D子组的包膜是相关的，并且B特异性引物还扩增了D亚组病毒。我们还设计了一套通用引物来扩增任何ALV亚组病毒。为了证明这些引物的实用性，我们从衣阿华州兽医生物学中心获得了被ALV感染的鸡胚成纤维细胞上清液，该病毒感染了两种商业马立克氏病疫苗中的一种污染物。使用我们的PCR引物，我们证明污染物是A ALV亚组。我们克隆并测序了一部分包膜基因，并确认ALV是A亚型病毒。与典型的A亚型病毒不同，污染物ALV在细胞培养中的生长非常缓慢。我们还从污染病毒中克隆并测序了部分长末端重复序列（LTR）。发现该LTR与在内源性ALV（E亚组）中发现的LTR相似，并且与通常在A亚组病毒中正常发现的LTR相似。类似E的LTR可能解释了为什么污染物在细胞培养中生长得如此差。

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