摘要
利用新兴的环介导等温扩增技术,建立了一种灵敏、特异、快速的体外环介导等温扩增检测方法。针对鹅细小病毒VP3基因的8个位点设计了6条特异性引物,建立了一种快速检测鹅细小病毒的LAMP方法。试验结果表明LAMP方法在恒温65℃水浴下50min内即可完成GPV的检测,且其检测灵敏度达150pg.μL-1;与其他常见的病毒无交叉反应。以钙黄绿素和Mn2+作为荧光指示剂,可快速、直观判定反应结果,可见该方法为适合基层及现场快速检测、实用灵敏的分子生物学方法。
A novel speed LAMP assay for GPV was explored with Loop-mediated amplification （LAMP）. With two pairs of identify six positions in VP3 gene, a LAMP assay was developed for the rapid detection GPV. The results indicated that GPV was detected by LAMP assay at 65 C as early as 50 min of incubation in a simple water bath. The detection limit of the LAMP assay was 150 pg ~ /~L ~ of GPV genome. And no amplification was found with the samples of other viruses.＂ A mixture of calcein and Mn2＋ was used as fluorescent reagent to make the result of LAMP visible. Therefore, the simple and fast method is a potential useful technique for GPV detection in the field.
出处
《福建农业学报》
CAS
2013年第1期9-13,共5页
Fujian Journal of Agricultural Sciences
基金
国家公益性行业(农业)科研专项(201003012)
