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

Masahiro Sakai, Dolores Baxa, Tomofumi Kurobe, Tomoya Kono, Raghunath B. Shivappa, Ronald Hedrick

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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 °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).

Original languageEnglish (US)
Pages (from-to)27-31
Number of pages5
JournalAquaculture
Volume288
Issue number1-2
DOIs
StatePublished - Mar 2 2009

Fingerprint

Oncorhynchus clarkii
Oncorhynchus
rainbow
Oncorhynchus mykiss
amplification
Salmonidae
parasite
parasites
Microsporidia
polymerase chain reaction
RNA
detection limit
ribosomal RNA
DNA
loop-mediated isothermal amplification
detection
methodology
infection
temperature
method

Keywords

  • Detection
  • LAMP
  • Nucleospora salmonis
  • PCR
  • Rainbow trout

ASJC Scopus subject areas

  • Aquatic Science

Cite this

Detection of Nucleospora salmonis in cutthroat trout (Oncorhynchus clarki) and rainbow trout (Oncorhynchus mykiss) by loop-mediated isothermal amplification. / Sakai, Masahiro; Baxa, Dolores; Kurobe, Tomofumi; Kono, Tomoya; Shivappa, Raghunath B.; Hedrick, Ronald.

In: Aquaculture, Vol. 288, No. 1-2, 02.03.2009, p. 27-31.

Research output: Contribution to journalArticle

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