Development of a loop-mediated isothermal amplification assay for rapid detection of Burkholderia mallei

被引：13
作者：
Mirzai, S. [1 ]
Safi, S. [2 ]
Mossavari, N. [3 ]
Afshar, D. [4 ]
Bolourchian, M. [5 ]
机构：
[1] Islamic Azad Univ, Fac Specialized Vet Sci, Sci & Res Branch, Grad Vet Med, Tehran, Iran
[2] Islamic Azad Univ, Fac Specialized Vet Sci, Sci & Res Branch, Dept Pathol & Clin Pathol, Poonak Sq, Tehran 1477893855, Iran
[3] Razi Vaccine & Serum Res Inst, PPD Tuberculin Dept, Karaj, Iran
[4] Zanjan Univ Med Sci, Dept Microbiol, Zanjan, Iran
[5] Islamic Azad Univ, Fac Specialized Vet Sci, Sci & Res Branch, Vet Med, Tehran, Iran
来源：
CELLULAR AND MOLECULAR BIOLOGY | 2016年 / 62卷 / 10期
关键词：
Glanders; identification; Burkholderia mallei; LAMP; LAMP ASSAY; PSEUDOMALLEI; VIRUS; IDENTIFICATION; ESTABLISHMENT; DIAGNOSIS; GLANDERS; SYSTEM; AGENT;
D O I：
10.14715/cmb/2016.62.10.5
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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 63 degrees C 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.
引用
收藏
页码：32 / 36
页数：5
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