摘要
本研究利用鸡W染色体中EE0.6序列上的Eco R片段为靶序列,进行引物设计,建立了基于LAMP法的快速鸟类性别鉴定方法。试验表明,该引物可用于多属种鸟类的性别鉴定,在64℃条件下,60 min即可完成检测。同时利用实时浊度法和荧光目视法两种LAMP检测法对体系进行了敏感性试验,并与PCR方法进行比较,实时浊度法与荧光目视法敏感性相近,是PCR方法的100倍。方法建立后,在大连老虎滩鸟语林进行了11目14科35种鸟类的性别鉴定临床试验,符合率为100%。
In this study,a rapid test assay was established based on Loop-Mediated Isothermal Amplification（LAMP）and the primers were designed by targeting EE0.6 pseudogene sequence on W chromosome from chicken.The reaction condition of the detection system was optimized and the results showed that the optimal condition was at 64 ℃ for 60 min.The assay can be available to various species of birds.The minimum detection limit of two types of LAMP assay（real-time turbidmeter,fluorescence visual method）were similar,and approximately 100-fold more sensitive than that of PCR.35 species of birds samples were collected from 11 orders and 14 families at Da Lian Laohutan Ocean Park Niaoyu Lin and tested by LAMP assay.The test accuracy rates was 100%.
出处
《现代畜牧兽医》
2017年第3期12-18,共7页
Modern Journal of Animal Husbandry and Veterinary Medicine
关键词
LAMP
鸟类
性别鉴定
实时浊度法
荧光目视法
LAMP
Bird
Sex identification
Real-time turbidmeter
Fluorescence visual method
