Development and validation of an RNA- and DNA-based quantitative PCR assay for determination of Kudoa thyrsites infection levels in Atlantic salmon Salmo salar

Diseases of Aquatic Organisms

Kristina M. Miller¹  Monique Raap¹  Simon Jones¹  Valerie A. Funk¹  Christy Falkenberg¹  John Robinson¹  Ken Sojonky¹ 

关键词: Kudoa thyrsites;    Atlantic salmon;    Diagnostics;    QPCR;    RNA test;    DNA test;    Histology;    Myoliquefaction;   

DOI  :  10.3354/dao075239

学科分类：生物科学（综合）

来源: Inter-Research

【 摘 要 】

ABSTRACT: Quantitative PCR (QPCR) methods targeting the 18S rDNA gene (DNA QPCR) and cathepsin L mRNA (RNA QPCR) from Kudoa thyrsites (Gilchrist) were developed and compared with histology for determination of K. thyrsites infection levels in Atlantic salmon Salmo salar L. Both QPCR tests were specific, reproducible and sensitive down to 3 copies. DNA QPCR was able to detect lower K. thyrsites infection levels than those detected by RNA QPCR and histology. The higher sensitivity of the DNA-based test compared with the RNA-based test appeared to be biological in nature and suggested that when infection levels were low, there were fewer copies of cathepsin L mRNA than 18S rDNA genes. However, all 3 diagnostic methods were highly correlated. Regression analyses comparing DNA QPCR and histology data from 2 distinct groups of fish showed that the relationship between these 2 diagnostic methods was reproducible. A logistic regression analysis comparing diagnostic data with a visual assessment of post-mortem flesh quality indicated that histology was the single best predictor of flesh quality, followed by DNA QPCR and then RNA QPCR.

【 授权许可】

Unknown