OIE white spot syndrome virus PCR gives false-positive results in Cherax quadricarinatus

Diseases of Aquatic Organisms

Bradford Cullen¹  Kerry Claydon¹  Leigh Owens¹ 

关键词: Cherax quadricarinatus;    Intranuclear bacilliform virus;    White spot syndrome virus;    PCR;   

DOI  :  10.3354/dao062265

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: White spot syndrome virus (WSSV) is an intranuclear bacilliform virus (IBV) that is a serious, notifiable crustacean pathogen. The Office International des Epizooties (OIE) PCR protocol for WSSV uses primer sets initially developed by Lo et al.(1996). It yields a first-step PCR amplicon of 1441 bp and a nested PCR amplicon of 941 bp. An amplicon (941 bp) purported to specifically detect WSSV was obtained when using template DNA extracted from Cherax quadricarinatus in a WSSV PCRdetection protocol recommended by the OIE. Sequencing and analysis of the 941 bp amplicon and an occasional 550 bp amplicon from C. quadricarinatus revealed no phylogenetic relationship with WSSV, and suggested a possible lack of sufficient primerspecificity for WSSV in the OIE test. This suggestion was supported by the fact that the OIE outer primer sequence (146F1) was present in both the forward and reverse position of the 941 bp and the forward position of the 550 bp nested amplicons fromC. quadricarinatus. As WSSV is a notifiable pathogen, the consequences of false-positive results are harsh in WSSV-free zones and can lead to incorrect quarantine and unnecessary destruction of animals. Therefore, urgent attention and revision isnecessary for the current OIE PCR protocol for WSSV detection.

【 授权许可】

Unknown