Development of a loop-mediated isothermal amplification assay for rapid detection of Burkholderia mallei.
The present study was conducted to establish a Loop-mediated isothermal amplification (LAMP) technique for the rapid detection of B. mallei the etiologic agent of glanders, a highly contagious disease of equines. A set of six specific primers targeting integrase gene cluster were designed for the LAMP test. The reaction was optimized using different temperatures and time intervals. The specificity of the assay was evaluated using DNA from B.pseudomallei and Pseudomonas aeruginosa. The LAMP products were analyzed both visually and under UV light after electrophoresis. The optimized conditions were found to be at 63ºC for 60 min. The assay showed high specificity and sensitivity. It was concluded that the established LAMP assay is a rapid, sensitive and practical tool for detection of B. mallei and early diagnosis of glanders.
Islamic Azad University Graduate of Veterinary Medicine, Faculty of Specialized Veterinary Sciences, Science and Research Branch Tehran Iran.
S
S
Mirzai
Islamic Azad University Department of Pathology and Clinical pathology, Faculty of Specialized Veterinary Sciences, Science and Research Branch Tehran Iran safishahab@yahoo.com.
S
S
Safi
Razi Vaccine & Serum Research Institute PPD Tuberculin Department Karaj Iran.
N
N
Mossavari
Zanjan University of Medical Sciences Department of Microbiology Zanjan Iran.
D
D
Afshar
Islamic Azad University Student of Veterinary Medicine, Faculty of Specialized Veterinary Sciences, Science and Research Branch Tehran Iran.
M
M
Bolourchian
D000818:Animals; D042726:Burkholderia mallei / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005453:Fluorescence; D006736:Horses; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction
D004269:DNA, Bacterial