Development of cross-priming amplification assays for rapid and sensitive detection of Aeromonas hydrophila

Abstract

Aeromonas hydrophila 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 Aer. hydrophila, 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 Aer. hydrophila. This novel method could be practical for use in primary care facilities and smaller clinical laboratories.

Development of a real‐time recombinase polymerase amplification assay for rapid detection of Aeromonas hydrophila

“…However, software for RPA primer design is not currently available but the design of the RPA primers and probes is key factor in optimizing RPA performance. The other molecular biological detections for A. hydrophila included real-time PCR (Griffin et al, 2013), LAMP (Chang et al, 2013;He et al, 2010) and CPA (Meng et al, 2015). We met the limit of detections of the CPA assay at 200 copies per reaction.…”

“…However, the primer design for LAMP techniques is complex and requires a specific, long and highly conserved DNA fragment that has limited its use for pathogen detection (Manmohan et al, 2008). Cross-priming amplification (CPA) is a recent isothermal DNA amplification technology but requires multiple cross-linked primers and complex reaction components (Meng et al, 2015;Zhong & Zhao, 2018). Therefore, a portable and user-friendly method is therefore necessary for the early point-ofneed detection of A. hydrophila infections.…”

Emerging Biorecognition and Transduction Schemes for Rapid Detection of Pathogenic Bacteria in Food

“…This advanced PCR technique is costly, but has one of the lowest LOD values in the literature for a culture‐independent technique associated with viability testing. This method has great promise, and use of viability indicators such as sigDE operon are under investigation by a number of labs to refine the method (Barnett and others ; Shao and others ; Ruff and others ; Garrido and others ; Garrido‐Maestu and others ; Li and others ; Vondrakova and others ; Zhou and others ; Meng and others ; Sebastião and others ). The drawback of this technique is the need for continuous monitoring of viability markers, complicating the assay, and adding analysis time and adding additional cost.…”

Sensitive Detection of Aeromonas Hydrophila by Time-Resolved Fluorimmunoassay

“…嗜水气单胞菌(Aeromonas hydrophila)属于弧菌科、 气单胞菌属, 它不仅引发人类败血症、脑膜炎、腹泻等 疾病 [1] , 而且还导致哺乳类等多种动物全身性败血症或 局部感染 [2] . 嗜水气单胞菌也是水产养殖业的重要病原 菌之一, 爆发时会引起水产经济动物的大面积死亡 [3,4] , 据报道, 鳗鲡败血病、红鳍病、烂鳃病均与嗜水气单胞 菌感染有关 [5] , 建立灵敏、特异、快速的检测技术, 对嗜 水气单胞菌进行监测, 不仅可减小病害造成的损失, 而 且可对疾病的预防和控制起到积极的指导作用.…”