Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, 250 Road Kuo Kuang, National Chung Hsing University, Taichung 402, Taiwan.
J Virol Methods. 2012 Apr;181(1):117-24. doi: 10.1016/j.jviromet.2011.09.002. Epub 2011 Sep 7.
Infectious bursal disease (IBD), an immunosuppressive disease that affects all ages of chickens, results in significant losses in the poultry industry. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with a chromatographic lateral flow dipstick (LFD) for the detection of infectious bursal disease virus (IBDV) was developed. The whole process of testing can be completed in less than 70 min using biotin-labeled primers, an FITC-labeled DNA probe, and the LFD. The detection limits for IBDV using RT-LAMP and RT-LAMP-LFD were the same at 10(-1)plaque forming units (PFU). When other unrelated viruses and cells were tested, no false positive results were observed. In addition, the amplification efficiency of RT-LAMP was enhanced when a loop primer was used. The RT-LAMP-LFD product started to be detected after 40 min. Clinical samples were used to compare assays using RT-PCR, nested RT-PCR, RT-LAMP, and RT-LAMP-LFD and the positive rates were 16%, 40%, 40%, and 40%, respectively. In conclusion, this assay is an easy, rapid, accurate, and sensitive method for the detection of IBDV and will improve the screening of field samples, especially when veterinarians have limited resources.
传染性腔上囊病（IBD）是一种影响所有年龄段鸡的免疫抑制性疾病，给家禽业造成了重大损失。本研究开发了一种用于检测传染性腔上囊病病毒（IBDV）的逆转录环介导等温扩增（RT-LAMP）联合层析侧向流试纸条（LFD）检测方法。使用生物素标记引物、FITC 标记 DNA 探针和 LFD，整个检测过程可在 70 min 内完成。使用 RT-LAMP 和 RT-LAMP-LFD 的 IBDV 检测限相同，均为 10(-1)噬斑形成单位（PFU）。当测试其他不相关的病毒和细胞时，没有观察到假阳性结果。此外，使用环引物增强了 RT-LAMP 的扩增效率。RT-LAMP-LFD 产物在 40 min 后开始检测。使用 RT-PCR、嵌套 RT-PCR、RT-LAMP 和 RT-LAMP-LFD 对临床样本进行比较检测，阳性率分别为 16%、40%、40%和 40%。综上所述，该检测方法是一种简便、快速、准确、灵敏的 IBDV 检测方法，将提高现场样本的筛选效率，特别是在兽医资源有限的情况下。
