Zeng Tingting, Xie Zhixun, Xie Liji, Deng Xianwen, Xie Zhiqin, Luo Sisi, Huang Li, Huang Jiaoling
Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
Virol J. 2015 Dec 30;12:226. doi: 10.1186/s12985-015-0455-5.
Immunosuppressive viruses are frequently found as co-infections in the chicken industry, potentially causing serious economic losses. Because traditional molecular biology methods have limited detection ability, a rapid, high-throughput method for the differential diagnosis of these viruses is needed. The objective of this study is to develop a GenomeLab Gene Expression Profiler Analyser-based multiplex PCR method (GeXP-multiplex PCR) for simultaneous detection of eight immunosuppressive chicken viruses.
Using chimeric primers, eight such viruses, including Marek's disease virus (MDV), three subgroups of avian leucosis virus (ALV-A/B/J), reticuloendotheliosis virus (REV), infectious bursal disease virus (IBDV), chicken infectious anaemia virus (CIAV) and avian reovirus (ARV), were amplified and identified by their respective amplicon sizes. The specificity and sensitivity of the optimised GeXP-multiplex PCR assay were evaluated, and the data demonstrated that this technique could selectively amplify these eight viruses at a sensitivity of 100 copies/20 μl when all eight viruses were present. Among 300 examined clinical specimens, 190 were found to be positive for immunosuppressive viruses according to this novel assay.
The GeXP-multiplex PCR assay is a high-throughput, sensitive and specific method for the detection of eight immunosuppressive viruses and can be used for differential diagnosis and molecular epidemiological surveys.
免疫抑制性病毒在养鸡业中常作为共感染出现，可能造成严重的经济损失。由于传统分子生物学方法的检测能力有限，因此需要一种快速、高通量的方法来对这些病毒进行鉴别诊断。本研究的目的是开发一种基于基因组实验室基因表达谱分析仪的多重PCR方法（GeXP-多重PCR），用于同时检测八种鸡免疫抑制性病毒。
使用嵌合引物，扩增并通过各自扩增子大小鉴定了八种此类病毒，包括马立克氏病病毒（MDV）、禽白血病病毒三个亚群（ALV-A/B/J）、网状内皮组织增生症病毒（REV）、传染性法氏囊病病毒（IBDV）、鸡传染性贫血病毒（CIAV）和禽呼肠孤病毒（ARV）。对优化后的GeXP-多重PCR检测方法的特异性和灵敏度进行了评估，数据表明，当所有八种病毒都存在时，该技术能够以100拷贝/20μl的灵敏度选择性地扩增这八种病毒。在300份检测的临床样本中，根据这种新检测方法，发现190份样本的免疫抑制性病毒呈阳性。
GeXP-多重PCR检测方法是一种用于检测八种免疫抑制性病毒的高通量、灵敏且特异的方法，可用于鉴别诊断和分子流行病学调查。
