Development of cross‐priming amplification assays for rapid and sensitive detection of Aeromonas hydrophila. (15th June 2015)

• Record Type: Journal Article
• Authors: Meng, S.; Wang, Y.; Wang, Y.; Liu, D.; Ye, C.
• Is Part Of: Letters in applied microbiology. Volume 61:Number 2(2015:Aug.)
• Page Start: 171
• Page End: 178
• Publication Date: 2015-06-15
• Subjects: Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.12439
• Languages: English
• ISSNs: 0266-8254

Abstract

<abstract abstract-type="main" id="lam12439-abs-0001"> <title>Abstract</title> <sec id="lam12439-sec-1001" sec-type="section"> <p> <italic>Aeromonas hydrophila</italic> has been increasingly implicated as the aetiologic agent of various human diseases. Therefore, reliable laboratory detection and identification of this bacterium has become clinically and epidemiologically desirable. We developed a nearly instrument‐free, simple molecular method for rapid detection of <italic>Aer. hydrophila</italic> using a cross‐priming amplification (CPA) assay with the <italic>desA</italic> gene as the target. The <italic>desA</italic> gene is crucial for the survival and growth of <italic>Aer. hydrophila</italic> under iron starvation. The results can be visualized as colour changes without opening the reaction tubes. No false‐positive results were observed for the 33 non‐<italic>Aer. hydrophila</italic> strains tested to evaluate assay specificity. The limit of detection for <italic>Aer. hydrophila</italic> was approximately 200 copies of <italic>desA</italic> per reaction (on reference plasmids) and 5 × 10<sup>3</sup> CFU g<sup>−1</sup><italic>Aer. hydrophila</italic> in simulated human stool, which is the same sensitivity as a qPCR assay. The performance of the CPA assay was also evaluated with 100 stool specimens from diarrhoea patients and 40 environmental water samples. In conclusion, the simplicity, cost‐effectiveness and nearly instrument‐free platform of the CPA assay make it practical for use in primary care facilities and smaller clinical laboratories.</p> </sec> <sec id="lam12439-sec-1002" sec-type="section"> <title>Significance and Impact of the Study</title> <p> <italic>Aeromonas hydrophila</italic> is a human pathogen that infects via exposed wounds or ingestion of contaminated water and food. In this study, a CPA‐based PCR method was developed for specific, rapid, cost‐effective detection of <italic>Aer. hydrophila</italic>, and the test results could be visualized without opening the reaction tubes. This is the first report on the application of the CPA method for the detection of <italic>Aer. hydrophila</italic>. This novel method could be practical for use in primary care facilities and smaller clinical laboratories.</p> </sec> </abstract>