Single and nested polymerase chain reaction assays for the detection of Microsporidium seriolae (Microspora), the causative agent of 'Beko' disease in yellowtail Seriola quinqueradiata

Citation
As. Bell et al., Single and nested polymerase chain reaction assays for the detection of Microsporidium seriolae (Microspora), the causative agent of 'Beko' disease in yellowtail Seriola quinqueradiata, DIS AQU ORG, 37(2), 1999, pp. 127-134
Citations number
23
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Aquatic Sciences
Journal title
DISEASES OF AQUATIC ORGANISMS
ISSN journal
0177-5103 → ACNP
Volume
37
Issue
2
Year of publication
1999
Pages
127 - 134
Database
ISI
SICI code
0177-5103(19990730)37:2<127:SANPCR>2.0.ZU;2-G
Abstract
Single and nested polymerase chain reaction (PCR) assays were developed for the detection of the microsporidian parasite Microsporidium seriolae, whic h is responsible for emaciation and even death in farmed Japanese yellowtai l. Extremely high rDNA identities exist between this parasite and other mem bers of the as yet unclassified genus, necessitating the design of generic, rather than species-specific primer sets. The nested PCR was several order s of magnitude more sensitive than the standard single PCRs, with visible t arget product amplified from as little as 0.01 pg of parasite DNA (equivale nt to that extracted from a single spore). The specificity of the assays wa s tested against a range of potential host fishes and 6 other microsporidia ns infecting either fish or the musculature of their hosts. Single PCRs wer e found to be specific to the target genus, but the nested PCR replicated r DNA from several different microsporidian genera, limiting its utility. Thi s study highlights problems associated with the use of the rRNA gene for PC R assays of certain microsporidians, but nevertheless provides a rapid and sensitive means for the detection of pre-spore forms not possible by curren t staining methods. Consequently, these assays may be employed for further studies on the portals of entry, migration to the musculature and transmiss ion of this economically important pathogen.