Identification of Vibrio anguillarum in fish by using partial 16S rRNA sequences and a specific 16S rRNA oligonucleotide probe.
16S rRNA from seven different Vibrio anguillarum strains was partially sequenced and compared. From this sequence information we could design a 25-base-long oligonucleotide and use it as a specific probe for identification of V. anguillarum. This was determined by RNA-DNA colony hybridization and slot-blot hybridization. Strong, specific hybridization to the probe was observed for all V. anguillarum strains tested. Furthermore, no cross-hybridization could be seen against five other bacterial species. The detection limit was 5 x 10(3) bacteria per ml. It was even possible to detect V. anguillarum, by slot-blot hybridization, directly in a homogenized kidney from a fish that had died of vibriosis. The partial sequence information revealed small but significant differences between strains of the same species. These sequence differences are sufficiently significant to allow serotyping on the RNA level. Comparing strains of different serotypes revealed a 10-base and an 11-base difference in V. anguillarum serotypes O8 and O9, respectively, in a 122-base partial sequence.
Department of Microbiology, University of Umeå, Sweden.
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Rehnstam
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Norqvist
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Wolf-Watz
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Hagström
D000818:Animals; D005393:Fish Diseases / Q000382:microbiology*; D005399:Fishes; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D013045:Species Specificity; D014337:Trout; D014733:Vibrio / Q000235:genetics / Q000302:isolation & purification*; D014735:Vibrio Infections / Q000382:microbiology / Q000662:veterinary*
D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S