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白斑综合征病毒(WSSV)聚合酶链反应(PCR)阳性的卤虫卵孵化出的无节幼体PCR检测呈阴性,将这些无节幼体投喂给对虾幼体时,无法传播WSSV。

White spot syndrome virus (WSSV) PCR-positive Artemia cysts yield PCR-negative nauplii that fail to transmit WSSV when fed to shrimp postlarvae.

作者信息

Chang Yun-Shiang, Lo Chu-Fang, Peng Shao-En, Liu Kuan-Fu, Wang Chung-Hsiung, Kou Guang-Hsiung

机构信息

Department of Zoology, National Taiwan University, Taipei, ROC.

出版信息

Dis Aquat Organ. 2002 Apr 24;49(1):1-10. doi: 10.3354/dao049001.

Abstract

Positive results were obtained with nested white spot syndrome virus (WSSV) diagnostic PCR performed on 5 commercial brands of dry-packed Artemia cysts using several WSSV genomic sequence-specific primers. In 2 brands, PCR and nucleotide sequence analysis found C-->T and C-->G point mutations in the pms 146 WSSV amplicon, but in all 5 brands, the nucleotide sequences that were successfully amplified by the rrl, rr2 and tk-tmk gene-specific primer sets were identical to those of Penaeus monodon WSSV. However, despite the inarguable presence of WSSV or WSSV-like template DNA, we were unable to detect WSSV by PCR in hatched nauplii derived from PCR-positive cysts or in P. monodon postlarvae fed Artemia nauplii hatched from such cysts. Most simply, these results suggested that the cysts were externally contaminated with WSSV or WSSV-like template material that was removed during hatching and washing of the nauplii. Given the small sequence variations found, it may also have been a variety of WSSV non-infectious for P. monodon or Artemia and derived from other crustaceans or arthropods in the Artemia environment. However, we could not establish this conclusively and a small possibility remained that the PCR template in these tests was derived from WSSV template present internally in the cysts and derived from infected Artemia adults. However small, this possibility must be vigorously tested, given the impact that a positive outcome could have on the shrimp industry.

摘要

使用几种白斑综合征病毒(WSSV)基因组序列特异性引物,对5个商业品牌的干包装卤虫囊肿进行巢式WSSV诊断PCR,获得了阳性结果。在2个品牌中,PCR和核苷酸序列分析发现pms 146 WSSV扩增子中存在C→T和C→G点突变,但在所有5个品牌中,由rrl、rr2和tk-tmk基因特异性引物组成功扩增的核苷酸序列与斑节对虾WSSV的核苷酸序列相同。然而,尽管无可争议地存在WSSV或WSSV样模板DNA,但我们无法通过PCR在源自PCR阳性囊肿的孵化无节幼体中或在喂食从此类囊肿孵化出的卤虫无节幼体的斑节对虾后期幼体中检测到WSSV。最简单地说,这些结果表明囊肿在外部被WSSV或WSSV样模板物质污染,这些物质在无节幼体孵化和清洗过程中被去除。鉴于发现的小序列变异,它也可能是对斑节对虾或卤虫无传染性的多种WSSV,源自卤虫环境中的其他甲壳类动物或节肢动物。然而,我们无法最终确定这一点,并且仍然存在一种小可能性,即这些测试中的PCR模板源自囊肿内部存在的WSSV模板,该模板源自受感染的卤虫成虫。鉴于阳性结果可能对虾产业产生的影响,无论可能性多小,都必须对这种可能性进行严格测试。

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