Rapid detection of Pseudomonas aeruginosa by cross priming amplification
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 dev...
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doaj-32b1edff1e9c46639f1bcf9d2c468f662021-06-08T04:42:27ZengElsevierJournal of Integrative Agriculture2095-31192020-10-01191025232529Rapid detection of Pseudomonas aeruginosa by cross priming amplificationYong XIANG0Ling YAN1Xiao-cui ZHENG2Li-zhen LI3Peng LIU4Wei-sheng CAO5College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, South China Agricultural University, Guangzhou 510642, P.R.China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, South China Agricultural University, Guangzhou 510642, P.R.China; 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; Correspondence CAO Wei-sheng, Tel: +86-20-85282536, Mobile: +86-13318865369Pseudomonas 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.http://www.sciencedirect.com/science/article/pii/S2095311920631872Pseudomonas aeruginosacross priming amplificationisothermal amplificationrapid detectiondetection method |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yong XIANG Ling YAN Xiao-cui ZHENG Li-zhen LI Peng LIU Wei-sheng CAO |
spellingShingle |
Yong XIANG Ling YAN Xiao-cui ZHENG Li-zhen LI Peng LIU Wei-sheng CAO Rapid detection of Pseudomonas aeruginosa by cross priming amplification Journal of Integrative Agriculture Pseudomonas aeruginosa cross priming amplification isothermal amplification rapid detection detection method |
author_facet |
Yong XIANG Ling YAN Xiao-cui ZHENG Li-zhen LI Peng LIU Wei-sheng CAO |
author_sort |
Yong XIANG |
title |
Rapid detection of Pseudomonas aeruginosa by cross priming amplification |
title_short |
Rapid detection of Pseudomonas aeruginosa by cross priming amplification |
title_full |
Rapid detection of Pseudomonas aeruginosa by cross priming amplification |
title_fullStr |
Rapid detection of Pseudomonas aeruginosa by cross priming amplification |
title_full_unstemmed |
Rapid detection of Pseudomonas aeruginosa by cross priming amplification |
title_sort |
rapid detection of pseudomonas aeruginosa by cross priming amplification |
publisher |
Elsevier |
series |
Journal of Integrative Agriculture |
issn |
2095-3119 |
publishDate |
2020-10-01 |
description |
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. |
topic |
Pseudomonas aeruginosa cross priming amplification isothermal amplification rapid detection detection method |
url |
http://www.sciencedirect.com/science/article/pii/S2095311920631872 |
work_keys_str_mv |
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