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<oai_dc:dc xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
  <dc:contributor>Rachel L. Powers</dc:contributor>
  <dc:contributor>Bonnie Besijn</dc:contributor>
  <dc:contributor>Paul K. Hershberger</dc:contributor>
  <dc:creator>Maureen K. Purcell</dc:creator>
  <dc:date>2017</dc:date>
  <dc:description>&lt;p&gt;&lt;span&gt;We report the development and validation of two quantitative PCR (qPCR) assays to detect&amp;nbsp;&lt;/span&gt;&lt;i&gt;Nanophyetus salmincola&lt;/i&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;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&lt;span&gt;&amp;nbsp;&lt;/span&gt;&lt;/span&gt;&lt;i&gt;N. salmincola&lt;/i&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;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&lt;span&gt;&amp;nbsp;&lt;/span&gt;&lt;/span&gt;&lt;i&gt;N. salmincola&lt;/i&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;DNA in settled water samples. Artificially spiked samples suggested that the 1-cercaria/L threshold corresponded to an estimated log&lt;/span&gt;&lt;sub&gt;10&lt;/sub&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;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&lt;span&gt;&amp;nbsp;&lt;/span&gt;&lt;/span&gt;&lt;i&gt;N. salmincola&lt;/i&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;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&lt;span&gt;&amp;nbsp;&lt;/span&gt;&lt;/span&gt;&lt;i&gt;N. salmincola&lt;/i&gt;&lt;span&gt;&lt;span&gt;&amp;nbsp;&lt;/span&gt;that occur in fish tissues, snail tissues, and water.&lt;/span&gt;&lt;/p&gt;</dc:description>
  <dc:format>application/pdf</dc:format>
  <dc:identifier>10.1080/08997659.2017.1365780</dc:identifier>
  <dc:language>en</dc:language>
  <dc:publisher>Taylor &amp; Francis</dc:publisher>
  <dc:title>Detection of Nanophyetus salmincola in water, snails, and fish tissues by quantitative polymerase chain reaction</dc:title>
  <dc:type>article</dc:type>
</oai_dc:dc>