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ID 25653
file
creator
Genmoto, K.
Nishizawa, Toyohiko
Muroga, Kiyokuni
subject
Vibrio penaeicida
rRNA
PCR Kuruma prawn
Vibriosis
NDC
Zoology
abstract
Vibrio penaeicida is the causative bacterium of vibriosis in cultured kuruma prawn Penaeus japonicus in Japan. To develop a specific and sensitive method for the detection of the pathogen, a species-specific sequence in the 16S rRNA of V. penaeicida was determined and a polymerase chain reaction (PCR)-based method was devised on the basis of the sequence. Prior to sequencing, a part of the variable regions of the 16S rRNA was amplified by using primers designed from 2 conserved regions according to previously reported data on Vibrionaceae. The region of the 16S rRNA (nucleotide numbers 440 to 490 in Escherichia coli 16S rRNA) obtained by this procedure was found to be species-specific for V. penaeicida. It was confirmed that PCR and RT (reverse transcription)-PCR amplifications with a sense primer designed from the V. penaeicida-specific sequence were both able to differentiate V. penaeicida from other prawn-pathogenic vibrios. 16S rRNA-targeted RT-PCR was demonstrated to have 100 times higher sensitivity than 16S rDNA-targeted PCR and 10 fg of total nucleic acids extracted from cultured bacterial cells was sufficient to yield the visible fragment in gel electrophoresis. These results indicate that RT-PCR amplification with this primer is useful for specific and sensitive detection of V. penaeicida.
journal title
Diseases of Aquatic Organisms
volume
Volume 24
issue
Issue 3
start page
185
end page
189
date of issued
1996-03-07
publisher
Inter-Research
issn
0177-5103
1616-1580
ncid
publisher doi
language
eng
nii type
Journal Article
HU type
Journal Articles
DCMI type
text
format
application/pdf
text version
publisher
rights
Copyright (c) 1996 Inter-Research.
relation url
department
Graduate School of Biosphere Science



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