Development of an Absolute Quantitative Real-Time PCR (qPCR) for the Diagnosis of
Aeromonas hydrophila
Infections in Fish
Fernanda de Alexandre Sebastião1, Eliana Gertrudes de Macedo Lemos2 and Fabiana Pilarski3*
1Department of Veterinary Medicine and Epidemiology, University of California, United States of America
2Department of Technology, Univ Estadual Paulista, Brazil
3Aquaculture Center of São Paulo State University (CAUNESP), Univ Estadual Paulista (UNESP), Brazil
*Corresponding Author: Fabiana Pilarski, Aquaculture Center of São Paulo State University (CAUNESP), Univ Estadual Paulista (UN
-
ESP), Brazil.
Received:
January 29, 2018; Published: March 14, 2018
DOI: 10.31080/ASMI.2018.01.0034
Aeromonas hydrophila
is a Gram-negative motile bacillus responsible for substantial losses in global aquaculture, which also poses
a risk to human health. Aeromonosis outbreaks are associated with excess organic matter in water supplies, failure in management,
primary parasitic infestations, and the presence of powerful bacterial virulence factors. This study validated an adhesin gene from
A.
hydrophila
(
ahaI
) using a quantitative real-time PCR assay based on SYBR Green I. The analytical specificity and sensitivity of the test
was analyzed. The ability to identify and quantify
A. hydrophila
samples isolated using classical microbiology techniques and total
DNA extracted directly from Nile tilapia,
Oreochromis niloticus
, kidney tissue experimentally infected with bacteria was also deter-
mined. The assay had 100% specificity and the sensitivity was 1.9 log gene copies of detection for total DNA from tissue. This study
constitutes a valuable tool for the rapid and accurate diagnosis of
A. hydrophila
in intensive fish farming scenarios.
Keywords: Adhesin Gene; Molecular Diagnosis; Nile Tilapia; Septicemia
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