Meng S, Wang Y, Wang Y, Liu D, Ye C
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, China CDC, Beijing, China.
Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China.
Lett Appl Microbiol. 2015 Aug;61(2):171-8. doi: 10.1111/lam.12439. Epub 2015 Jun 15.
Aeromonas hydrophila 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 Aer. hydrophila using a cross-priming amplification (CPA) assay with the desA gene as the target. The desA gene is crucial for the survival and growth of Aer. hydrophila 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-Aer. hydrophila strains tested to evaluate assay specificity. The limit of detection for Aer. hydrophila was approximately 200 copies of desA per reaction (on reference plasmids) and 5 × 10(3) CFU g(-1) Aer. hydrophila 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.
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.
嗜水气单胞菌越来越多地被认为是各种人类疾病的病原体。因此，可靠地在实验室检测和鉴定这种细菌在临床和流行病学方面都很有必要。我们开发了一种几乎无需仪器的简单分子方法，以desA基因为靶点，通过交叉引物扩增（CPA）检测法快速检测嗜水气单胞菌。desA基因对于嗜水气单胞菌在铁饥饿条件下的存活和生长至关重要。结果可通过颜色变化直观呈现，无需打开反应管。对33株非嗜水气单胞菌菌株进行检测以评估检测特异性，未观察到假阳性结果。嗜水气单胞菌的检测限约为每个反应200拷贝的desA（基于参考质粒）以及模拟人粪便中5×10³CFU g⁻¹嗜水气单胞菌，这与定量聚合酶链反应（qPCR）检测法的灵敏度相同。还使用100份腹泻患者的粪便标本和40份环境水样评估了CPA检测法的性能。总之，CPA检测法的简单性、成本效益和几乎无需仪器的平台使其适用于基层医疗设施和较小的临床实验室。
嗜水气单胞菌是一种通过暴露伤口或摄入受污染的水和食物进行感染的人类病原体。在本研究中，开发了一种基于CPA的聚合酶链反应方法，用于特异性、快速、经济高效地检测嗜水气单胞菌，且检测结果无需打开反应管即可直观呈现。这是关于应用CPA方法检测嗜水气单胞菌的首次报道。这种新方法可能适用于基层医疗设施和较小的临床实验室。
