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

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. Gol...

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Published in:PloS one 2018-10, Vol.13 (10), p.e0204332-e0204332
Main Authors: Niu, Lina, Zhao, Fan, Chen, Jinlong, Nong, Jinqing, Wang, Chunmei, Wang, Jing, Gao, Naishu, Zhu, Xiaoxue, Wu, Lei, Hu, Shoukui
Format: Article
Language:English
Subjects:
Amplification
Assaying
Biology and Life Sciences
Biosensing Techniques - methods
Biosensors
Colorimetry
Deoxyribonucleic acid
Design
DNA
Engineering and Technology
Genes
Gold
Gold - chemistry
Hospitalization
Humans
Klebsiella
Klebsiella pneumoniae
Klebsiella pneumoniae - genetics
Klebsiella pneumoniae - isolation & purification
Laboratories
Limit of Detection
Medicine and Health Sciences
Metal Nanoparticles - chemistry
Methods
Nanoparticles
Nosocomial infection
Nosocomial infections
Nucleic Acid Amplification Techniques - methods
Outbreaks
Pneumonia
Polymerase chain reaction
Research and Analysis Methods
Sputum
Staphylococcus infections
Temperature
Time Factors
Tuberculosis
Turbidimeters
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cited_by cdi_FETCH-LOGICAL-c526t-56ed20a23bd7f370959fd9a65a1fa4ea87912b65647b24b37b908341644f83783
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creator Niu, Lina
Zhao, Fan
Chen, Jinlong
Nong, Jinqing
Wang, Chunmei
Wang, Jing
Gao, Naishu
Zhu, Xiaoxue
Wu, Lei
Hu, Shoukui
description 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.
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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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>30273362</pmid><doi>10.1371/journal.pone.0204332</doi><orcidid>https://orcid.org/0000-0001-6339-3954</orcidid><oa>free_for_read</oa></addata></record>
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subjects Amplification
Assaying
Biology and Life Sciences
Biosensing Techniques - methods
Biosensors
Colorimetry
Deoxyribonucleic acid
Design
DNA
Engineering and Technology
Genes
Gold
Gold - chemistry
Hospitalization
Humans
Klebsiella
Klebsiella pneumoniae
Klebsiella pneumoniae - genetics
Klebsiella pneumoniae - isolation & purification
Laboratories
Limit of Detection
Medicine and Health Sciences
Metal Nanoparticles - chemistry
Methods
Nanoparticles
Nosocomial infection
Nosocomial infections
Nucleic Acid Amplification Techniques - methods
Outbreaks
Pneumonia
Polymerase chain reaction
Research and Analysis Methods
Sputum
Staphylococcus infections
Temperature
Time Factors
Tuberculosis
Turbidimeters
title Isothermal amplification and rapid detection of Klebsiella pneumoniae based on the multiple cross displacement amplification (MCDA) and gold nanoparticle lateral flow biosensor (LFB)
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