Rapid and Sensitive Detection of Vibrio alginolyticus by Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Dipstick Targeted to the rpoX Gene
Vibrio alginolyticus is a major bacterial pathogen causing disease in marine animals. The present study aimed to develop a loop-mediated isothermal amplification (LAMP) coupled with a lateral flow dipstick (LFD) for rapid and simple visual detection of V. alginolyticus-specific amplicons. The biotin...
| Veröffentlicht in: | Journal of aquatic animal health. - 1998. - 27(2015), 3 vom: 28. Sept., Seite 156-63 |
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| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2015
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| Zugriff auf das übergeordnete Werk: | Journal of aquatic animal health |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't Bacterial Proteins Molecular Probes |
| Zusammenfassung: | Vibrio alginolyticus is a major bacterial pathogen causing disease in marine animals. The present study aimed to develop a loop-mediated isothermal amplification (LAMP) coupled with a lateral flow dipstick (LFD) for rapid and simple visual detection of V. alginolyticus-specific amplicons. The biotin-labeled LAMP amplicons from the targeted portion of a gene encoding rpoS-like sigma factor (rpoX) were generated at 60°C for 1 h and then hybridized with a fluorescein isothiocyanate-labeled probe for 5 min for visual detection with LFD. In pure cultures, the detection limit of the LAMP-LFD technique for V. alginolyticus was 1.8 × 10(2) CFU/mL while that of PCR was 1.8 × 10(3) CFU/mL. In spiked whiteleg shrimp samples Penaeus vannamei, the sensitivity for V. alginolyticus detection was 2 × 10(3) CFU/g (equivalent to 4 CFU per reaction) while PCR was 10 times less sensitive. The LAMP-LFD method for V. alginolyticus correctly identified 21 isolates of V. alginolyticus but did not recognize 23 non-V. alginolyticus Vibrio isolates and 15 non-Vibrio isolates. In summary, this LAMP-LFD method targeted to the rpoX gene is a convenient assay for specific identification of V. alginolyticus with high sensitivity |
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| Beschreibung: | Date Completed 30.11.2015 Date Revised 30.09.2020 published: Print Citation Status MEDLINE |
| ISSN: | 1548-8667 |
| DOI: | 10.1080/08997659.2015.1037468 |