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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Differential Detection of Avian Oncogenic Viruses in Poultry Layer Farms and Turkeys by Use of Multiplex PCR
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Differential Detection of Avian Oncogenic Viruses in Poultry Layer Farms and Turkeys by Use of Multiplex PCR

机译：使用多重PCR差异检测禽肉场农场和土耳其的禽致癌病毒

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Avian oncogenic viruses include Marek's disease virus (MDV), a highly contagious herpesvirus, as well as retroviruses such as avian leukosis virus (ALV) subgroups A to J and reticuloendotheliosis virus (REV). In this study, we examined the incidence of these viruses in suspected samples collected from poultry layer farms of South India, mainly in the Namakkal district of Tamil Nadu, a highly dense poultry-growing area in India. The histopathology-positive tissue sections were identified and further confirmed by immunohistochemistry using virus-specific antibodies. The viruses belonging to all 3 groups (MDV, ALV, and REV) were isolated in a cell culture system and confirmed by immunofluorescence using virus-specific antibodies. PCR appeared to be the method of choice for rapid and accurate diagnosis of these viruses. The multiplex PCR primers specific to MDV, ALV, REV, and chicken DNA were designed for rapid differential diagnosis. The specificity of the primers was checked by amplification of DNA from virus-infected cell culture in comparison with uninfected samples, and sensitivity was evaluated by calculating the minimum copy number at which amplification occurs in the cloned PCR products. The sequences of the amplicons were compared by BLAST analysis. PCR tests demonstrated the presence of single, dual, or triple viruses in some of the samples. Of 169 samples screened by multiplex PCR, 9 samples were positive for MDV, 17 samples were positive for ALV, 12 samples were positive for REV, and 17 samples were positive for both ALV and REV. Three samples were positive for all three viruses. ALV-positive samples were further subjected to subgroup-specific PCR, which gave positive results for subgroups B and D but not for subgroup J. Multiplex PCR appeared to be a useful technique for rapid differential diagnosis of avian oncogenic viruses and detection of multiple infections of avian oncogenic viruses under field conditions.
机译：禽致癌病毒包括高度传染性疱疹病毒马立克氏病病毒（MDV），以及禽白血病病毒（ALV）A至J亚组和网状内皮内皮病病毒（REV）等逆转录病毒。在这项研究中，我们检查了从印度南部（主要是印度高密度禽类生长区泰米尔纳德邦）的纳马克卡勒地区的禽类农场收集的可疑样本中这些病毒的发生率。鉴定组织病理学阳性组织切片，并使用病毒特异性抗体通过免疫组织化学进一步确认。在细胞培养系统中分离出属于所有3组（MDV，ALV和REV）的病毒，并使用病毒特异性抗体通过免疫荧光进行确认。 PCR似乎是快速准确诊断这些病毒的首选方法。设计用于MDV，ALV，REV和鸡DNA的多重PCR引物，用于快速鉴别诊断。与未感染的样品相比，通过从病毒感染的细胞培养物中扩增DNA来检查引物的特异性，并通过计算在克隆的PCR产物中发生扩增的最小拷贝数来评估敏感性。通过BLAST分析比较扩增子的序列。 PCR测试表明某些样品中存在单，双或三重病毒。多重PCR筛选的169个样品中，MDV阳性9个，ALV阳性17个，REV阳性12个，ALV和REV阳性17个。对于所有三种病毒，三个样本均为阳性。对ALV阳性样品进一步进行亚组特异性PCR，该分析对于B和D组给出了阳性结果，但对于J组却没有给出阳性结果。多重PCR似乎是快速鉴别禽类致癌病毒和检测多种感染的有用技术。野外条件下的禽致癌病毒。

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