Chongqing Academy of Animal Science, Chongqing 402460, Chongqing, China.
Virol J. 2010 Jan 21;7:14. doi: 10.1186/1743-422X-7-14.
Goose parvovirus (GPV) is a Dependovirus associated with latent infection and mortality in geese. Currently, in a worldwide scale, GPV severely affects geese production. The objective of this study is to develop a loop-mediated isothermal amplification (LAMP) method for the sensitive, rapid, and inexpensive detection of GPV in the field.
A set of six specific primers was designed by targeting the GPV VP3 DNA. With Bst DNA polymerase large fragment, the target DNA could be amplified at 65 degrees C as early as 20 min of incubation in a simple water bath. A positive reaction was identified through the detection of the LAMP product by color change visible to the naked eye. The detection limit of the assay was 28 copies/microl of plasmid pVP3, and with equal sensitivity and specificity to fluorescent quantitative real-time PCR (FQ-PCR).
The high sensitivity, specificity, and simplicity, as well as the high throughput, make this method suitable for specific detection of GPV infection in both field conditions and laboratory settings. The utilization of complicated equipment and conduct of technical training on the GPV LAMP were not necessary.
鹅细小病毒(GPV)是一种依赖病毒,与鹅的潜伏感染和死亡率有关。目前,在全球范围内,GPV 严重影响鹅的生产。本研究旨在开发一种环介导等温扩增(LAMP)方法,用于在现场敏感、快速和廉价地检测 GPV。
针对 GPV VP3 DNA 设计了一组六个特异性引物。使用 Bst DNA 聚合酶大片段,在简单的水浴中孵育 20 分钟即可在 65°C 下扩增靶 DNA。通过肉眼可见的颜色变化检测 LAMP 产物来识别阳性反应。该测定的检测限为 28 拷贝/微升质粒 pVP3,与荧光定量实时 PCR(FQ-PCR)具有相同的灵敏度和特异性。
该方法具有高灵敏度、特异性和简单性,以及高通量,使其适用于现场和实验室条件下 GPV 感染的特异性检测。不需要使用复杂的设备和对 GPV LAMP 进行技术培训。