A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss)

被引：101

作者：

Gunimaladevi, I

Kono, T

LaPatra, SE

Sakai, M

机构：

[1] Miyazaki Univ, Fac Agr, Miyazaki 8892192, Japan

[2] Kagoshima Univ, United Grad Sch Agr Sci, Kagoshima 890, Japan

[3] Clear Springs Foods Inc, Div Res, Buhl, ID USA

来源：

ARCHIVES OF VIROLOGY | 2005年 / 150卷 / 05期

关键词：

D O I：

10.1007/s00705-004-0468-7

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout (Oncorhynchus mykiss). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 degrees C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a 10-fold higher sensitivity was seen for the LAMP protocols.

引用

页码：899 / 909

页数：11

共 49 条

[1] Nested PCR improves detection of infectious hematopoietic necrosis virus in cells coinfected with infectious pancreatic necrosis virus [J].