Detection of koi herpesvirus in common carp, Cyprinus carpio L., by loop-mediated isothermal amplification

被引:80
作者
Gunimaladevi, I
Kono, T
Venugopal, MN
Sakai, M
机构
[1] Miyazaki Univ, Fac Agr, Miyazaki 8892192, Japan
[2] Kagoshima Univ, United Grad Sch Agr Sci, Kagoshima 890, Japan
[3] Univ Agr Sci Mangalore, Coll Fisheries, Dept Fishery Microbiol, Mangalore 575002, India
关键词
detection; KHV Japanese strain; KHV-LAMP; KHV-PCR; koi herpesvirus; tk gene;
D O I
10.1111/j.1365-2761.2004.00578.x
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. In this study, using the LAMP method, a protocol for koi herpes virus (KHV) detection in common carp was designed. A set of four primers, two inner and two outer, were designed based on the sequence of the thymidine kinase (tk) gene of KHV. Time and temperature conditions for detection of KHV were optimized for 60 min at 65 degreesC. The detection limit using LAMP was found to be similar to that by polymerase chain reaction. In this study, we have developed a highly sensitive and rapid diagnostic procedure for detection of KHV infection in common carp.
引用
收藏
页码:583 / 589
页数:7
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