title: Development of a rapid and specific loop-mediated isothermal amplification detection method that targets Marek's disease virus meq gene.
abstract: A rapid, sensitive and specific loop-mediated isothermal amplification (LAMP) method was developed and evaluated for the detection of Marek's disease virus (MDV) by amplification of conserved MDV meq gene sequences. LAMP is an innovative technique that allows the rapid detection of targeted nucleic acid sequences under isothermal conditions without the need for complex instrumentation. In this study, meq gene sequences were amplified successfully from different MDV strains by LAMP within 60min and no cross-reactivity was observed in a panel of related viruses that were associated with diseases of chickens. The detection limit of LAMP was 3.2 copies/million cells compared with 320 copies/million cells required for conventional PCR. Positive detection rates were assessed using either LAMP or PCR by examination of feather follicles that were collected from chickens infected experimentally with either strain J-1 (n=20) or strain Md5 (n=17), In addition to these samples, three isolates that were suspected to have been infected in the clinic were also tested. Results showed that the positive detection rate for LAMP was 95% (38/40), compared with 87.5% (35/40) and 90% (38/40) for strains J-1 and Md5 by PCR, respectively. These results indicated that the LAMP assay was more sensitive, rapid and specific than conventional PCR for the detection of MDV. This easy-to-perform technique will be useful for the detection of MDV and will aid in the establishment of disease control protocols.
authors.affiliation: Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Xiuying
authors.initials: X
authors.lastname: Wei
authors.forename: Xingming
authors.initials: X
authors.lastname: Shi
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Mei
authors.initials: M
authors.lastname: Wang
authors.forename: Changjun
authors.initials: C
authors.lastname: Liu
authors.forename: Hongyu
authors.initials: H
authors.lastname: Cui
authors.forename: Shunlei
authors.initials: S
authors.lastname: Hu
authors.forename: Yanming
authors.initials: Y
authors.lastname: Quan
authors.forename: Hongyan
authors.initials: H
authors.lastname: Chen
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002645:Chickens / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005814:Genes, Viral; D025961:Herpesvirus 3, Gallid / Q000235:genetics* / Q000302:isolation & purification; D057230:Limit of Detection; D008380:Marek Disease / Q000175:diagnosis / Q000821:virology*; D025202:Molecular Diagnostic Techniques; D021141:Nucleic Acid Amplification Techniques
chemical_list: D004279:DNA, Viral