Simultaneous detection of eight immunosuppressive chicken viruses using a GeXP analyser-based multiplex PCR assay
- PDF / 1,289,629 Bytes
- 12 Pages / 595.276 x 790.866 pts Page_size
- 68 Downloads / 200 Views
METHODOLOGY
Open Access
Simultaneous detection of eight immunosuppressive chicken viruses using a GeXP analyser-based multiplex PCR assay Tingting Zeng, Zhixun Xie*, Liji Xie, Xianwen Deng, Zhiqin Xie, Sisi Luo, Li Huang and Jiaoling Huang
Abstract Background: 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. Results: 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. Conclusion: 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. Keywords: GeXP analyser, Multiplex PCR, Immunosuppressive viruses
Background Immunosuppression causes major economic losses in poultry farming because immunosuppressed chickens are more susceptible to viral and bacterial pathogens, respond poorly to vaccination, and display lower feed conversion efficiency as well as growth retardation. Immunosuppressive chicken viruses include Marek’s disease virus (MDV), avian leucosis virus (ALV), reticuloendotheliosis virus (REV), infectious bursal disease virus (IBDV), chicken infectious anaemia virus (CIAV) and avian reovirus (ARV), all of which affect immune function in chickens and lead to immunosuppression [1–7]. The typical symptoms elicited by these viruses differ, and some symptoms are not readily observable. * Correspondence: [email protected] Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi 530001, China
Immunosuppression in chickens infected with MDV, ALV or REV occurs much earlier than does tumour development and death [1, 2]. Although early infection with IBDV in chicks less than 3 weeks old may not result in the typical symptoms of IBD, this infection nonetheless causes serious immunosuppression [8]. Chickens older than 3 weeks of age are resistant to anaemia after infec
Data Loading...
