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Journal of Integrative Agriculture  2020, Vol. 19 Issue (10): 2523-2529    DOI: 10.1016/S2095-3119(20)63187-2
Special Issue: 动物医学合辑Veterninary Medicine
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
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 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
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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.
Keywords:  Pseudomonas aeruginosa        cross priming amplification        isothermal amplification        rapid detection        detection method  
Received: 22 October 2019   Accepted:
Fund: 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).
Corresponding Authors:  Correspondence CAO Wei-sheng, Tel: +86-20-85282536, Mobile: +86-13318865369, E-mail:   

Cite this article: 

XIANG Yong, YAN Ling, ZHENG Xiao-cui, LI Li-zhen, LIU Peng, CAO Wei-sheng. 2020. Rapid detection of Pseudomonas aeruginosa by cross priming amplification. Journal of Integrative Agriculture, 19(10): 2523-2529.

Abdouchakour F, Aujoulat F, Licznar-Fajardo P, Marchandin H, Toubiana M, Parer S. 2018. Intraclonal variations of resistance and phenotype in Pseudomonas aeruginosa epidemic high-risk clone ST308: A key to success within a hospital? International Journal of Medical Microbiology, 308, 279–289.
Agodi A, Barchitta M, Cipresso R, Giaquinta L, Romeo M A, Denaro C. 2007. Pseudomonas aeruginosa carriage, colonization, and infection in ICU patients. Intensive Care Medicine, 33, 1155–1161.
Amoureux L, Riedweg K, Chapuis A, Bador J, Siebor E, Pechinot A. 2017. Nosocomial infections with IMP-19-producing Pseudomonas aeruginosa linked to contaminated sinks, France. Emerging Infectious Diseases, 23, 304–307.
Devriese L A, Viaene N J, Demedts G. 1975. Pseudomonas aeruginosa infection on a broiler farm. Avian Pathology, 4, 233–237.
Fang R, Li X, Hu L, You Q, Li J, Wu J. 2009. Cross-priming amplification for rapid detection of Mycobacterium tuberculosis in sputum specimens. Journal of Clinical Microbiology, 47, 845–847.
Feng T, Li S, Wang S, Pan J. 2018. Cross priming amplification with nucleic acid test strip analysis of mutton in meat mixtures. Food Chemistry, 245, 641–645.
Fernandes M R, Sellera F P, Moura Q, Carvalho M P N, Rosato P N, Cerdeira L. 2018. Zooanthroponotic transmission of drug-resistant Pseudomonas aeruginosa, Brazil. Emerging Infectious Diseases, 24, 1160–1162.
Gao Y, Meng X Y, Zhang H W, Luo Y Z, Sun Y, Li Y F. 2018. Cross-priming amplification combined with immunochromatographic strip for rapid on-site detection of African swine fever virus. Sensors and Actuators (B: Chemical), 274, 304–309.
Gou H, Li J, Cai R, Song S, Li M, Yang D. 2018. Rapid detection of Haemophilus parasuis using cross-priming amplification and vertical flow visualization. Journal of Microbiological Methods, 144, 67–72.
Karampatakis T, Antachopoulos C, Tsakris A, Roilides E. 2018. Molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa in an endemic area: Comparison with global data. European Journal of Clinical Microbiology and Infectious Diseases, 37, 1211–1220.
Kidd T J, Soares Magalhaes R J, Paynter S, Bell S C, Group A C I. 2015. The social network of cystic fibrosis centre care and shared Pseudomonas aeruginosa strain infection: A cross-sectional analysis. The Lancet Respiratory Medicine, 3, 640–650.
Kuboki N, Inoue N, Sakurai T, Di Cello F, Grab D J, Suzuki H. 2003. Loop-mediated isothermal amplification for detection of African trypanosomes. Journal of Clinical Microbiology, 41, 5517–5524.
Lai H, Zhang H, Ning Z, Chen R, Zhang W, Qing A. 2011. Isolation and characterization of emerging subgroup J avian leukosis virus associated with hemangioma in egg-type chickens. Veterinary Microbiology, 151, 275–283.
Oliver A, Mulet X, Lopez-Causape C, Juan C. 2015. The increasing threat of Pseudomonas aeruginosa high-risk clones. Drug Resistance Updates, 21–22, 41–59.
Parida M, Shukla J, Sharma S, Santhosh R S, Ravi V, Mani R. 2011. Development and evaluation of reverse transcription loop-mediated isothermalamplification assay for rapid and real-time detection of the swine-origin influenza A H1N1 virus. The Journal of Molecular Diagnostics, 13, 100–107.
Saeed M, Rasheed F, Afzal R K, Hussain S, Riaz S, Ahmad A. 2018. Pseudomonas aeruginosa: Evaluation of pathogen burden and drug-resistance trends in a tertiary care hospital. Journal of the College of Physicians and Surgeons Pakistan, 28, 279–283.
Walker S E, Sander J E, Cline J L, Helton J S. 2002. Characterization of Pseudomonas aeruginosa isolates associated with mortality in broiler chicks. Avian Diseases, 46, 1045–1050.
Wang Y X, Zhang A Y, Yang Y Q, Lei C W, Cheng G Y, Zou W C. 2018. Sensitive and rapid detection of Salmonella enterica serovar Indiana by cross-priming amplification. Journal of Microbiological Methods, 153, 24–30.
Xu G, Hu L, Zhong H, Wang H, Yusa S, Weiss T C. 2012. Cross priming amplification: Mechanism and optimization for isothermal DNA amplification. Scientific Reports, 2, 246.
Yang H L, Huang J, Yang B, Liu F, Zhang Q L. 2014. The establishment and application of isothermal cross-priming amplification techniques in detecting penaeid shrimp white spot syndrome virus. Letters in Applied Microbiology, 59, 200–206.
Zhang J, Liu X, Li W, Zhang J, Xiao Z, Zhou Z. 2018. Rapid detection of milk vetch dwarf virus by loop-mediated isothermal amplification. Journal of Virological Methods, 261, 147–152.
Zhang R, Liu Z, Li J, Lei L, Yin W, Li M. 2017. Presence of VIM-positive Pseudomonas species in chickens and their surrounding environment. Antimicrob Agents Chemother, 61, e00167–e00183.
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