鲤浮肿病检测方法实验室间比对的结果与评价

Inter-laboratory comparison results and evaluation of different carp edema disease detection methods

鲤浮肿病(carp edema virus disease,CEVD)是在锦鲤中发现的一种传染性病毒病,近年来在全球传播迅速,对鲤养殖业造成严重损害。本研究设计CEV环介导等温扩增(loop-mediated isothermal amplification,LAMP)引物,进行特异性实验;选取套式PCR、荧光PCR和LAMP三种检测方法,组织10家实验室进行室间比对测试:制备含CEV病毒核酸的测试样品,评价其均匀性和稳定性,通过测试结果分析方法检出限。结果显示,套式PCR的核酸样品检测限可达6.8×10^(-10) g。

Carp edema virus disease ( CEVD) is an infectious virus disease found in koi carp. It has spreaded rapidly around the world in recent years, and causing great economic losses to koi and common carp breeding industry. In this study, CEV loop-mediated isothermal amplification ( LAMP) primers were designed to carry out specific tests. Three detection methods including nested PCR, fluorescent PCR and lamp were selected and 10 laboratories were organized to carry out inter-laboratory comparison. Test samples containing CEV virus nucleic acid were prepared, and their uniformity and stability were evaluated. The results showed that the detection limit of nucleic acid samples by nested PCR could reach from 6.8×10 -10 g. A rapid on-site detection method for CEV was established in this study.