Detection of Nucleospora salmonis in cutthroat trout (Oncorhynchus clarki) and rainbow trout (Oncorhynchus mykiss) by loop-mediated isothermal amplification

被引：7

作者：

Sakai, Masahiro ^{[1
]}

Baxa, Dolores V. ^{[2
]}

Kurobe, Tomofumi ^{[2
]}

Kono, Tomoya ^{[1
]}

Shivappa, Raghunath B. ^{[3
]}

Hedrick, Ronald P. ^{[2
]}

机构：

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

[2] Univ Calif Davis, Sch Vet Med, Dept Med & Epidemiol, Davis, CA 95616 USA

[3] N Carolina State Univ, Coll Vet Med, Raleigh, NC 27606 USA

来源：

AQUACULTURE | 2009年 / 288卷 / 1-2期

关键词：

Nucleospora salmonis; Detection; PCR; LAMP; Rainbow trout; POLYMERASE-CHAIN-REACTION; CHINOOK SALMON; INTRANUCLEAR MICROSPORIDIUM; RAPID DETECTION; FISH; INFECTIONS; HEDRICK; DNA;

D O I：

10.1016/j.aquaculture.2008.11.005

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

Nucleospora salmonis is an intranuclear microsporidian parasite of salmonids that causes a chronic lymphoblastosis and a leukemic-like condition in both cultured and wild salmonids. Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. Using the LAMP method, a protocol for detection of N. salmonis was developed. A set of four primers, two inner and two outer were designed based on the sequence of ribosomal RNA intermediate spacer of this parasite. Optimum time and temperature conditions for detection of N. salmonis by LAMP were 60 min at 63 degrees C, respectively. The detection-limit (DL) using LAMP was found to be similar to polymerase chain reaction (PCR). In this study, we have developed a highly sensitive and rapid diagnostic procedure for detection of N. salmonis infections in cutthroat trout(Oncorhynchus clarki) and rainbow trout (Oncorhynchus mykiss). (C) 2008 Elsevier B.V. All rights reserved,

引用

页码：27 / 31

页数：5

共 22 条

[1] NESTED POLYMERASE CHAIN-REACTION FOR DETECTION OF ENTEROCYTOZOON SALMONIS GENOMIC DNA IN CHINOOK SALMON ONCORHYNCHUS-TSHAWYTSCHA [J].