A sensitive FRET probe assay for the selective detection of Mycobacterium marinum in fish

Journal of Fish Diseases
Fulvio SalatiGermano Orrù

Abstract

Mycobacterium marinum is the causative agent of mycobacteriosis in wild and cultured fish and of atypical infection in humans. For the diagnosis of M. marinum, cultural and traditional polymerase chain reaction (PCR) methods are currently used. However, these protocols, although able to discriminate within Mycobacterium spp., have proved to be time-consuming or difficult to carry out. For this reason, the aim of this study was to obtain a rapid and specific diagnostic tool to quantify fish Mycobacterium spp. or to discriminate M. marinum from other mycobacteria. A primary PCR amplification with SYBR Green had a detection limit (dl) of 10(2)Mycobacterium DNA copies with a log-linear quantification range up to 10(4) (R(2) = 0.99). The second PCR using FRET probes, flanking a region containing species specific nucleotide variations, was designed and validated with synthetic erp gene fragments corresponding to different mycobacterial species, different whole mycobacteria suspensions, experimentally infected fish tissues, tissues from experimentally infected fish, and samples of cultured fish. The results show that the FRET probes demonstrate a high specificity as the melting curve analysis allowed efficient discrimination of M. mar...Continue Reading

References

Oct 1, 1992·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·J A Vazquez, J D Sobel
Sep 1, 1992·American Journal of Medical Genetics·P S Davies, C Joughin
Apr 1, 1992·International Journal of Systematic Bacteriology·V V Lévy-Frébault, F Portaels
Sep 1, 1989·Oral Microbiology and Immunology·G L SmithS S Socransky
Sep 1, 1993·Molecular Phylogenetics and Evolution·H SchneiderM Goodman
Aug 31, 2002·Journal of Clinical Microbiology·Jacqueline LachnikFranz-Christoph Bange
Oct 31, 2002·Applied and Environmental Microbiology·M UckoW R Knibb
Dec 21, 2002·European Journal of Cardio-thoracic Surgery : Official Journal of the European Association for Cardio-thoracic Surgery·Sevval ErenOmer Satici
Dec 9, 2004·Diseases of Aquatic Organisms·Martha W RhodesChristopher A Ottinger
Feb 8, 2005·Journal of Clinical Microbiology·M Ucko, A Colorni
Apr 9, 2005·Journal of Clinical Microbiology·Anthony AblordeyPhilip Supply
Jun 16, 2005·Journal of Clinical Microbiology·Aine SkowKaren L Kaul
Jan 3, 2006·Journal of Medical Microbiology·M Hui, W T Kwok
Mar 24, 2006·FEMS Microbiology Letters·Michael L KentLuiz E Bermudez
Apr 15, 2006·The Cancer Journal·Catherine Van Poznak
Sep 5, 2006·Journal of Fish Diseases·I M KaattariE B Shotts
May 8, 2007·Emerging Infectious Diseases·Andrew S KaneAna M Baya
May 26, 2007·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·Lesla S Bruijnesteijn van CoppenraetEdward J Kuijper

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Citations

Apr 20, 2011·The Journal of Parasitology·Mauro MeloniFulvio Salati
Oct 15, 2013·Applied and Environmental Microbiology·Thu Nguyet PhungJean-Christophe Avarre
Jan 7, 2014·BioMed Research International·Michal SlanyMonika Bodnarova
Apr 20, 2013·Veterinary Parasitology·Fulvio SalatiGiulia Angelucci
Oct 6, 2012·Journal of Molecular Microbiology and Biotechnology·Michal Slany, Ivo Pavlik
Mar 16, 2018·Clinical Practice and Epidemiology in Mental Health : CP & EMH·Germano Orrù, Mauro Giovanni Carta
Mar 16, 2018·Clinical Practice and Epidemiology in Mental Health : CP & EMH·Germano OrrùAlessia Bramanti
Sep 11, 2020·Microorganisms·Mohammad Reza DelghandiSimon Menanteau-Ledouble
Apr 8, 2017·Microbiology Spectrum·Alexandra AubryEmmanuelle Cambau

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