Detection of Nanophyetus salmincola in water, snails, and fish tissues by quantitative polymerase chain reaction

Journal of Aquatic Animal Health
By: , and 

Links

Abstract

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.

Additional publication details

Publication type Article
Publication Subtype Journal Article
Title Detection of Nanophyetus salmincola in water, snails, and fish tissues by quantitative polymerase chain reaction
Series title Journal of Aquatic Animal Health
DOI 10.1080/08997659.2017.1365780
Volume 29
Issue 4
Year Published 2017
Language English
Publisher Taylor & Francis
Contributing office(s) Western Fisheries Research Center
Description 10 p.
First page 189
Last page 198