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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Development and Application of Real-Time PCR for Detection of Subgroup J Avian Leukosis Virus
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Development and Application of Real-Time PCR for Detection of Subgroup J Avian Leukosis Virus

机译：实时PCR检测亚组J禽白血病病毒的开发及应用

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摘要

Subgroup J avian leukosis virus (ALV-J) is an avian retrovirus that causes severe economic losses in the poultry industry. The early identification and removal of virus-shedding birds are important to reduce the spread of congenital and contact infections. In this study, a TaqMan-based real-time PCR method for the rapid detection and quantification of ALV-J with proviral DNA was developed. This method exhibited a high specificity for ALV-J. Moreover, the detection limit was as low as 10 viral DNA copies. The coefficients of variation (CVs) of both interassay and intra-assay reproducibility were less than 1%. The growth curves of ALV-J in DF-1 cells were measured by real-time PCR, yielding a trend line similar to those determined by 50% tissue culture infective dose (TCID50) and p27 antigen detection. Tissue samples suspected of ALV infection were evaluated using real-time PCR, virus isolation, and routine PCR, and the positivity rates were 60.1%, 41.6% and 44.5%, respectively. Our data indicated that the real-time PCR method provides a sensitive, specific, and reproducible diagnostic tool for the identification and quantification of ALV-J for clinical diagnosis and in laboratory research.
机译：J禽白血病病毒亚组（ALV-J）是一种禽逆转录病毒，在禽类行业造成严重的经济损失。尽早发现并清除掉病毒的禽鸟对于减少先天性和接触性感染的传播很重要。在这项研究中，开发了一种基于TaqMan的实时PCR方法，用于用原病毒DNA快速检测和定量ALV-J。该方法对ALV-J表现出高特异性。而且，检出限低至10个病毒DNA拷贝。批间和批内重复性的变异系数（CVs）均小于1％。通过实时PCR测量ALV-J在DF-1细胞中的生长曲线，得出的趋势线类似于通过50％组织培养物感染剂量（TCID50）和p27抗原检测确定的趋势线。使用实时荧光定量PCR，病毒分离和常规PCR对怀疑有ALV感染的组织样品进行评估，阳性率分别为60.1％，41.6％和44.5％。我们的数据表明，实时PCR方法为临床诊断和实验室研究中ALV-J的鉴定和定量提供了灵敏，特异性和可重现的诊断工具。

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