Isothermal amplification and rapid detection of Klebsiella pneumoniae based on the multiple cross displacement amplification (MCDA) and gold nanoparticle lateral flow biosensor (LFB)

Autor: Fan Zhao, Gao Naishu, Shoukui Hu, Lei Wu, Chunmei Wang, Xiaoxue Zhu, Jing Wang, Jinlong Chen, Jinqing Nong, Lina Niu
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Time Factors
Physiology
Klebsiella pneumoniae
Staphylococcus
Metal Nanoparticles
lcsh:Medicine
Artificial Gene Amplification and Extension
Biosensing Techniques
Pathology and Laboratory Medicine
medicine.disease_cause
Polymerase Chain Reaction
Klebsiella Pneumoniae
law.invention
Limit of Detection
law
Klebsiella
Medicine and Health Sciences
Staphylococcus Aureus
lcsh:Science
Polymerase chain reaction
Gel Electrophoresis
Multidisciplinary
biology
Chemistry
Temperature
Detectors
Bacterial Pathogens
Body Fluids
Bioassays and Physiological Analysis
Medical Microbiology
Staphylococcus aureus
Engineering and Technology
Pathogens
Anatomy
medicine.symptom
Nucleic Acid Amplification Techniques
Research Article
030106 microbiology
Loop-mediated isothermal amplification
Equipment
Research and Analysis Methods
Microbiology
Rapid detection
Electrophoretic Techniques
03 medical and health sciences
medicine
Humans
Molecular Biology Techniques
Microbial Pathogens
Molecular Biology
Colorimetric Assays
Detection limit
Bacteria
lcsh:R
Organisms
Sputum
Biology and Life Sciences
biology.organism_classification
Molecular biology
respiratory tract diseases
Mucus
Biosensors
lcsh:Q
Gold
Biochemical Analysis
Biosensor
Zdroj: PLoS ONE, Vol 13, Iss 10, p e0204332 (2018)
PLoS ONE
ISSN: 1932-6203
Popis: Klebsiella pneumoniae (K. pneumoniae) is a frequent pathogen causing nosocomial infections and outbreaks. We developed a multiple cross displacement amplification (MCDA) assay for the detection of K. pneumoniae, which can get the positive results within 40 minutes’ isothermal amplification. Gold-nanoparticle lateral flow biosensor (LFB) and colorimetric indicators were used for the rapid readouts of MCDA amplification. The detection limit of this assay was 100 fg per reaction at 65°C, which was confirmed to be the optimal amplification temperature according to the real time turbidimeters. For specificity, all of the 30 clinical-source K. pneumoniae strains were positive for the MCDA, and all of the non-K. pneumoniae strains belonging to 31 different species were negative for this MCDA assay. To evaluate the practical applicability of this method, we assessed its detection limit for K. pneumoniae strains in sputum samples (24 CFU per reaction), and DNA templates of 100 sputum samples further underwent the MCDA-LFB tests. All of the sputum samples being positive for K. pneumoniae (30/100) with the culture method were successfully identified with the MCDA assay, the detection power of which was higher than that of polymerase chain reaction (PCR) (25/100). Thus, the MCDA test for K. pneumoniae combined with the gold nanoparticle LFB as the results readout scheme, are simple, specific, and sensitive methods for the rapid diagnosis of K. pneumoniae in clinical samples.
Databáze: OpenAIRE
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