title: Rapid detection and identification of nontuberculous mycobacterial pathogens in fish by using high-resolution melting analysis.
abstract: Mycobacterial infections in fish are commonly referred to as piscine mycobacteriosis, irrespectively of the specific identity of the causal organism. They usually cause a chronic disease and sometimes may result in high mortalities and severe economic losses. Nearly 20 species of Mycobacterium have been reported to infect fish. Among them, Mycobacterium marinum, M. fortuitum, and M. chelonae are generally considered the major agents responsible for fish mycobacteriosis. As no quick and inexpensive diagnostic test exists, we tested the potential of high-resolution melting analysis (HRMA) to rapidly identify and differentiate several Mycobacterium species involved in fish infections. By analyzing both the melting temperature and melting profile of the 16S-23S rRNA internal transcribed spacer (ITS), we were able to discriminate 12 different species simultaneously. Sensitivity tests conducted on purified M. marinum and M. fortuitum DNA revealed a limit of detection of 10 genome equivalents per reaction. The primers used in this procedure did not lead to any amplification signal with 16 control non-Mycobacterium species, thereby demonstrating their specificity for the genus Mycobacterium.
authors.affiliation: Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam.
authors.forename: Thu Nguyet
authors.initials: TN
authors.lastname: Phung
authors.forename: Domenico
authors.initials: D
authors.lastname: Caruso
authors.forename: Sylvain
authors.initials: S
authors.lastname: Godreuil
authors.forename: Nicolas
authors.initials: N
authors.lastname: Keck
authors.forename: Tatiana
authors.initials: T
authors.lastname: Vallaeys
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Avarre
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021901:DNA, Intergenic / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000382:microbiology*; D005399:Fishes / Q000382:microbiology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D009165:Mycobacterium Infections, Nontuberculous / Q000382:microbiology / Q000662:veterinary*; D009170:Nontuberculous Mycobacteria / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D044366:Transition Temperature; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D004269:DNA, Bacterial; D021901:DNA, Intergenic