title: Simultaneous detection of eight immunosuppressive chicken viruses using a GeXP analyser-based multiplex PCR assay.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zeng
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China. xiezhixun@126.com.
authors.forename: Zhixun
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Liji
authors.initials: L
authors.lastname: Xie
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Xianwen
authors.initials: X
authors.lastname: Deng
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Zhiqin
authors.initials: Z
authors.lastname: Xie
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Sisi
authors.initials: S
authors.lastname: Luo
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.affiliation: Guangxi Key Laboratory of Animal Vaccines and Diagnostics, Guangxi Veterinary Research Institute, 51 Youai North Road, Nanning, Guangxi, 530001, China.
authors.forename: Jiaoling
authors.initials: J
authors.lastname: Huang
mesh_terms: D000818:Animals; D017727:Chicken anemia virus; D002645:Chickens; D004266:DNA Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D003937:Diagnosis, Differential; D057166:High-Throughput Screening Assays; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology*; D012327:RNA Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods; D014773:Virology / Q000379:methods*