Nemoto Manabu, Ohta Minoru, Tsujimura Koji, Bannai Hiroshi, Yamanaka Takashi, Kondo Takashi, Matsumura Tomio
Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400–4 Shiba, Shimotsuke, Tochigi 329–0412, Japan.
J Vet Med Sci. 2011 Sep;73(9):1225-7. doi: 10.1292/jvms.11-0065. Epub 2011 May 6.
We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.
我们评估了环介导等温扩增技术（LAMP）作为直接从鼻拭子中检测1型马疱疹病毒（EHV-1）DNA的一种方法。为了提高灵敏度，我们在原始LAMP程序中增加了一个对样品进行热处理的步骤。即使省略DNA提取步骤，经过热处理的LAMP检测法的检测限也比原始LAMP检测法灵敏10倍。此外，经过热处理的LAMP检测法比原始LAMP检测法以及使用临床样本的聚合酶链反应更灵敏。经过热处理的LAMP检测法易于操作，因此应该适用于临床实验室对EHV-1感染的诊断。
