用于快速简便诊断牛白血病病毒感染的荧光环介导等温扩增检测方法的开发。
Development of a fluorescent loop-mediated isothermal amplification assay for rapid and simple diagnosis of bovine leukemia virus infection.
作者信息
Nagao Konomu, Makino Ryohei, Apego Francis Victor, Mekata Hirohisa, Yamazaki Wataru
机构信息
Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuen Kibanadai-nishi, Miyazaki, Miyazaki 889-2192, Japan.
Center for Animal Disease Control, University of Miyazaki, 1-1 Gakuen Kibanadai-nishi, Miyazaki, Miyazaki 889-2192, Japan.
出版信息
J Vet Med Sci. 2019 May 31;81(5):787-792. doi: 10.1292/jvms.19-0009. Epub 2019 Mar 27.
Bovine leukemia virus (BLV) causes enzootic bovine leukosis (EBL), a condition that threatens the sustainability of the livestock industry. A fluorescent loop-mediated isothermal amplification (fLAMP) assay targeting BLV env sequences was developed and used to evaluate 100 bovine blood samples. Compared with a conventional real-time PCR (rPCR) assay, the fLAMP assay achieved 87.3% (62/71) sensitivity and 100% (29/29) specificity. The rPCR assay took 65 min, while the fLAMP assay took 8 min to 30 min from the beginning of DNA amplification to final judgement with a comparable limit of detection. The fLAMP is a potential tool for the rapid and simple diagnosis of BLV infection to supplement ELISA testing and can be used by local laboratories and slaughterhouses without special equipment.
牛白血病病毒（BLV）可引发地方流行性牛白血病（EBL），这种疾病对畜牧业的可持续发展构成威胁。一种针对BLV env序列的荧光环介导等温扩增（fLAMP）检测方法被开发出来，并用于评估100份牛血样本。与传统的实时聚合酶链反应（rPCR）检测方法相比，fLAMP检测方法的灵敏度达到87.3%（62/71），特异性达到100%（29/29）。rPCR检测需要65分钟，而fLAMP检测从DNA扩增开始到最终判定需要8分钟至30分钟，两者的检测限相当。fLAMP是一种用于快速、简单诊断BLV感染的潜在工具，可作为酶联免疫吸附测定（ELISA）检测的补充，并且当地实验室和屠宰场无需特殊设备即可使用。
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