Abstract

Avian oncogenic or tumor diseases are common in poultry industry causing significant economic loss. Marek’s disease (MD), avian leukosis (AL) and Reticuloendotheliosis (RE) are the three major viral oncogenic infections that are difficult to differentiate with gross lesions. Multiplex PCR for simultaneous detection and differentiation of these three viruses was developed and validated. The primers targeting the genes of pp38, pol and LTR for MDV, ALV and REV were designed to yield 206, 429, and 128 bp, respectively. The sensitivity of the PCR primers was checked with serial dilution of positive template DNA for each virus and found to be in the range of 10⁻⁵ to 10⁻⁷ of 1 µg/µl of initial template DNA. Out of 114 suspected tumor samples screened, 8 samples were positive for MDV, 13 samples were positive for ALV and 31 samples positive for REV. Five samples were positive for both MD and ALV; 3 samples were positive for MD and REV and 25 samples were positive for ALV and REV. Eight samples were positive for all three viruses. Multiplex PCR demonstrated to be a useful technique for simultaneous, rapid detection and differentiation of major tumor causing and immunosuppressive viral diseases of chicken.

摘要

禽类致癌或肿瘤疾病在家禽业中很常见，造成重大经济损失。马立克氏病（MD）、禽白血病（AL）和网状内皮增生症（RE）是难以与肉眼病变鉴别的三大病毒性致癌性感染。开发并验证了用于同时检测和区分这三种病毒的多重 PCR。针对 MDV、ALV 和 REV 的pp38、pol和LTR基因设计的引物分别产生 206、429 和 128 bp。通过连续稀释每种病毒的阳性模板 DNA 来检查 PCR 引物的灵敏度，发现在 10 ^-5到 10 ^{-7的范围内}1 µg/µl 初始模板 DNA。在筛查的 114 个疑似肿瘤样本中，8 个样本为 MDV 阳性，13 个样本为 ALV 阳性，31 个样本为 REV 阳性。五个样本对 MD 和 ALV 均呈阳性；3个样本MD和REV呈阳性，25个样本ALV和REV呈阳性。八个样本对所有三种病毒均呈阳性。多重 PCR 被证明是一种有用的技术，可用于同时、快速检测和区分鸡的主要致瘤性和免疫抑制性病毒病。