title: Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification.
abstract: Mycoplasma synoviae (MS) remains a serious concern in production of poultry and affects world production of chickens and turkeys. Loop-mediated isothermal amplification (LAMP) of DNA has been recently used for the identification of different economically important avian pathogens. The aim of this study was to develop LAMP for simple and inexpensive detection of MS strains in poultry using specifically designed primers targeting hemagglutin A (vlh) gene. The assay was conducted in a water bath for 1 h at 63 °C. The results were visualized after addition of SYBR Green(®) fluorescent dye. LAMP was specific exclusively for MS without cross-reactivity with other Mycoplasma species. The sensitivity of LAMP was determined as 10(-1) CFU/ml and was 1,000 times higher than MS-specific polymerase chain reaction. LAMP assay was conducted on 18 MS field strains to ensure its reliability and usefulness. This is the first report on LAMP development and application for the rapid detection of MS isolated from chickens. This simple method may be applied by diagnostic laboratories without access to expensive equipment.
authors.affiliation: Department of Poultry Diseases, National Veterinary Research Institute, Partyzantów 57 Avenue, 24-100, Puławy, Poland.
authors.forename: Olimpia
authors.initials: O
authors.lastname: Kursa
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Woźniakowski
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Tomczyk
authors.forename: Anna
authors.initials: A
authors.lastname: Sawicka
authors.forename: Zenon
authors.initials: Z
authors.lastname: Minta
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D037102:Lectins / Q000235:genetics; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045805:Mycoplasma synoviae / Q000235:genetics* / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D037102:Lectins; C406455:VlhA protein, Mycoplasma synoviae