title: Rapid detection of infectious laryngotracheitis virus isolates by loop-mediated isothermal amplification.
abstract: The objective of this study was to develop and evaluate a loop-mediated isothermal amplification (LAMP) method to detect infectious laryngotracheitis virus (ILTV) from commercial broiler and layer flocks in southern China. A set of six specific primers was designed to recognize six distinct genomic sequences of thymidine kinase (TK) from ILTV. The entire assay duration was recorded at 40 min under isothermal condition at 63.5 degrees C. The amplified products were analyzed by electrophoresis and visual judgment by the SYBR Green I dyeing. LAMP assay was 10-fold more sensitive than the routine PCR assay, with a detection limit of 46 copies per reaction. In detecting ILTV, the LAMP assay detected all 5 strains previously isolated, did not cross-react with other avian pathogens, and obtained a 100% sensitivity in 43 positive clinical samples with reference to virus isolation. Therefore, the LAMP assay may be a good alternative method for specific diagnosis of ILTV infection in primary care facilities, and in less well-equipped laboratories.
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou 510642, China. qmx@scau.edu.cn
authors.forename: Qing-mei
authors.initials: QM
authors.lastname: Xie
authors.forename: Jun
authors.initials: J
authors.lastname: Ji
authors.forename: Tristan Tyler
authors.initials: TT
authors.lastname: Pickens
authors.forename: Li-qin
authors.initials: LQ
authors.lastname: Du
authors.forename: Yong-chang
authors.initials: YC
authors.lastname: Cao
authors.forename: Hong-mei
authors.initials: HM
authors.lastname: Li
authors.forename: Lin-guo
authors.initials: LG
authors.lastname: Wang
authors.forename: Jing-yun
authors.initials: JY
authors.lastname: Ma
authors.forename: Ying-zuo
authors.initials: YZ
authors.lastname: Bi
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002645:Chickens; D002681:China; D019411:Clinical Laboratory Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D005456:Fluorescent Dyes; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D004279:DNA, Viral; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I