Journal of Integrative Agriculture ›› 2020, Vol. 19 ›› Issue (10): 2523-2529.DOI: 10.1016/S2095-3119(20)63187-2

所属专题: 动物医学合辑Veterninary Medicine

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  • 收稿日期:2019-10-22 出版日期:2020-10-01 发布日期:2020-09-08

Rapid detection of Pseudomonas aeruginosa by cross priming amplification

XIANG Yong1, YAN Ling1, ZHENG Xiao-cui1, LI Li-zhen1, LIU Peng1, CAO Wei-sheng1, 2, 3, 4
  

  1. 1 College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China
    2 Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, South China Agricultural University, Guangzhou 510642, P.R.China
    3 National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, South China Agricultural University, Guangzhou 510642, P.R.China
    4 South China Collaborative Innovation Centre for Prevention and Control of Poultry Infectious Diseases and Safety of Poultry Products, South China Agricultural University, Guangzhou 510642, P.R.China
  • Received:2019-10-22 Online:2020-10-01 Published:2020-09-08
  • Contact: Correspondence CAO Wei-sheng, Tel: +86-20-85282536, Mobile: +86-13318865369, E-mail: caoweish@scau.edu.cn
  • Supported by:
    This study was supported by the Guangdong Key S&T Program (2019B020217002) from the Department of Science and Technology of Guangdong Province, China, the Guangdong Poultry Industry Technology System, China (2019KJ128) and the earmarked fund for China Agriculture Research System (CARS-41-G16).

Abstract:

Pseudomonas aeruginosa (PA) is an opportunistic pathogen of humans and animals and a common source of nosocomial infections especially of the respiratory tract.  Pseudomonas aeruginosa is also a major bacterial disease of poultry and in particular, eggs and newly hatched chicks.  In this study, we developed a simple, accurate and rapid molecular detection method using cross priming amplification (CPA) with a nucleic acid test strip to detect P. aeruginosa.  The assay efficiently amplified the target gene within 45 min at 62°C only using a simple water bath.  The detection limit of the method was 1.18×102 copies μL–1 for plasmid DNA and 4.4 CFU mL–1 for bacteria in pure culture, and was 100 times more sensitive than conventional PCR.  We screened 83 clinical samples from yellow-feather broiler breeder chickens and hospitalized/treated dogs and cats using CPA, PCR and traditional culture methods.  The positive-sample ratios were 15.3% (13/83) by CPA, 13.3% (11/83) by PCR and 12.1% (10/83) by the culture method.  The established CPA method has significant advantages for detecting P. aeruginosa.  The method is easy to use and possesses high specificity and sensitivity without the requirements of complicated experimental equipment.  The PA-CPA assay is especially fit for outdoor and primary medical units and is an ideal system for the rapid detection and monitoring of P. aeruginosa.

Key words: Pseudomonas aeruginosa ,  cross priming amplification ,  isothermal amplification ,  rapid detection ,  detection method