Detection of Nanophyetus salmincola in Water, Snails, and Fish Tissues by Quantitative Polymerase Chain Reaction
We report the development and validation of two quantitative PCR (qPCR) assays to detect Nanophyetus salmincola DNA in water samples and in fish and snail tissues. Analytical and diagnostic validation demonstrated good sensitivity, specificity, and repeatability of both qPCR assays. The N. salmincol...
| Veröffentlicht in: | Journal of aquatic animal health. - 1998. - 29(2017), 4 vom: 14. Dez., Seite 189-198 |
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| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2017
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| Zugriff auf das übergeordnete Werk: | Journal of aquatic animal health |
| Schlagworte: | Journal Article Water 059QF0KO0R |
| Zusammenfassung: | We report the development and validation of two quantitative PCR (qPCR) assays to detect Nanophyetus salmincola DNA in water samples and in fish and snail tissues. Analytical and diagnostic validation demonstrated good sensitivity, specificity, and repeatability of both qPCR assays. The N. salmincola DNA copy number in kidney tissue was significantly correlated with metacercaria counts based on microscopy. Extraction methods were optimized for the sensitive qPCR detection of N. salmincola DNA in settled water samples. Artificially spiked samples suggested that the 1-cercaria/L threshold corresponded to an estimated log10 copies per liter ≥ 6.0. Significant correlation of DNA copy number per liter and microscopic counts indicated that the estimated qPCR copy number was a good predictor of the number of waterborne cercariae. However, the detection of real-world samples below the estimated 1-cercaria/L threshold suggests that the assays may also detect other N. salmincola life stages, nonintact cercariae, or free DNA that settles with the debris. In summary, the qPCR assays reported here are suitable for identifying and quantifying all life stages of N. salmincola that occur in fish tissues, snail tissues, and water. Received April 13, 2017; accepted August 6, 2017 |
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| Beschreibung: | Date Completed 11.01.2018 Date Revised 30.09.2020 published: Print Citation Status MEDLINE |
| ISSN: | 1548-8667 |
| DOI: | 10.1080/08997659.2017.1365780 |