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Detection and Identification of Fish‐Pathogenic Aphanomyces piscicida Using Polymerase Chain Reaction (PCR) with Species‐Specific Primers
Author(s) -
Phadee Panarat,
Kurata Osamu,
Hatai Kishio,
Hirono Ikuo,
Aoki Takashi
Publication year - 2004
Publication title -
journal of aquatic animal health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 52
eISSN - 1548-8667
pISSN - 0899-7659
DOI - 10.1577/h03-047.1
Subject(s) - biology , polymerase chain reaction , primer (cosmetics) , microbiology and biotechnology , saprolegnia , pathogen , fungus , fish <actinopterygii> , gene , genetics , botany , fishery , chemistry , organic chemistry
Aphanomyces piscicida is an important pathogen that plays a role in the morbidity and mortality of various fish species around the world. The poor quality of current identification techniques for this fungus led to the development of highly sensitive and specific polymerase chain reaction (PCR) techniques. Primers derived from the ITS1 and ITS2 regions of A. piscicida NJM 0204 were designed for the detection and identification of fish‐pathogenic A. piscicida , with the potential for the diagnosis of mycotic granulomatosis (MG). Polymerase chain reaction amplification showed that the primer set was specific only to fish‐pathogenic A. piscicida , not to non‐fish‐pathogenic Aphanomyces , Saprolegnia spp., Achlya spp., Dictyuchus spp., and Lagenidium spp. In addition, PCR revealed an improved sensitivity sufficient to detect A. piscicida in artificially infected goldfish Carassius auratus . Results demonstrated that the PCR method established in this study is effective for the detection and identification of A. piscicida with MG.