[1]李文杰,周国勤,朱菲莉,等.检测克氏原螯虾白斑综合征病毒(WSSV)的巢式PCR方法的建立与初步应用[J].南京师大学报(自然科学版),2009,32(02):98-102.
　Li Wenjie,Zhou Guoqin,Zhu Feili,et al.Development and Potential Application of a Nested PCR for the Detection of White Spot Syndrome Virus in Procambarus Clarkia[J].Journal of Nanjing Normal University(Natural Science Edition),2009,32(02):98-102.
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检测克氏原螯虾白斑综合征病毒(WSSV)的巢式PCR方法的建立与初步应用()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第32卷
期数:
2009年02期
页码:
98-102
栏目:
生命科学
出版日期:
2009-06-30

文章信息/Info

Title:
Development and Potential Application of a Nested PCR for the Detection of White Spot Syndrome Virus in Procambarus Clarkia
作者:
李文杰;周国勤;朱菲莉;王卫东;周国平;
南京市水生动物疫病预防控制中心, 南京市水产科学研究所, 江苏南京210036
Author(s):
Li Wenjie; Zhou Guoqin; Zhu Feili; Wang Weidong; Zhou Guoping
Nanjing Center for Control and Prevention of Aquatic Animal Infections Disease,Nanjing Aquatic Sciences Institute,Nanjing 210036,China
关键词:
白斑综合征病毒;  巢式PCR;  克氏原螯虾
Keywords:
wh ite spot syndrom e v irus;  nested PCR;  Procam barus c la rkii
分类号:
S945.4
摘要:
根据GenBank中已发表的3株WSSV的VP28基因序列,设计了4条特异性引物.通过筛选最佳反应条件和试剂工作浓度,建立了检测克氏原螯虾WSSV的巢式PCR方法.该方法第一轮扩增的敏感性是36 ng,第二轮扩增的敏感性是36 pg.在对9份克氏原螯虾临床病料的巢式PCR检测中,用引物W1和W2做第一轮扩增,未检测到阳性病例;用引物S1和S2做第二轮扩增,检测到一例WSSV阳性病例.以上结果表明:对于检测克氏原螯虾WSSV,巢式PCR较一步PCR具有更高的检测灵敏度和应用前景.
Abstract:
Four specific pr im er, W1, W2, S1 and S2, we re des igned according to theVP28 gene sequences o f 3 w hite spo t syndrom e v irus (WSSV ) strains depo sited in the GenBank. PCR de tecting the VP28 gene ofW SSV from Procam ba rus clark ii was estab lished after se lection o f the optim al reaction cond itions. Sensitiv ity of the first and second amp lifications by the nested PCR w as 36 ng and 36 pg, respectively. Am ong 9 c lin ical Procam barus c la rkii sam ples to be tested by nested PCR, therew as no positive case detec ted by the first PCR amp lifications w ith pr im ersW1 andW2, and oneW SSV positive case detec ted by the second PCR amp lificationsw ith prim ers S1 and S2. The results showed that the nested PCR established by th is study w as much mo re su itab le than the one step PCR for the detection ofWSSV in Procambarus clark ii.

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备注/Memo

备注/Memo:
基金项目: 南京市水产专项资金( 2008)资助项目.
通讯联系人: 周国勤, 高级工程师, 研究方向: 水生动物疫病防治. E-m ail:xlfish@ vip. s ina. com
更新日期/Last Update: 2013-04-23