Please use this identifier to cite or link to this item: https://ir.swu.ac.th/jspui/handle/123456789/13588
ชื่อเรื่อง: Development of uracil-DNA-glycosylase-supplemented loop-mediated isothermal amplification coupled with nanogold probe (UDG-LAMP-AuNP) for specific detection of Pseudomonas aeruginosa
ผู้แต่ง: Manajit O.
Longyant S.
Sithigorngul P.
Chaivisuthangkura P.
Keywords: DNA polymerase
gold nanoparticle
RNA 16S
thymidine triphosphate
uracil DNA glycosidase
gold
metal nanoparticle
RNA 16S
uracil DNA glycosidase
amplicon
Article
bacterium contamination
bacterium detection
bacterium identification
colony forming unit
colorimetry
controlled study
cross reaction
DNA hybridization
ecfx gene
gene
limit of detection
loop mediated isothermal amplification
nonhuman
nucleic acid amplification
Pseudomonas aeruginosa
sensitivity and specificity
genetics
human
microbiology
molecular probe
polymerase chain reaction
procedures
Pseudomonas aeruginosa
Pseudomonas infection
Gold
Humans
Metal Nanoparticles
Molecular Probes
Nucleic Acid Amplification Techniques
Polymerase Chain Reaction
Pseudomonas aeruginosa
Pseudomonas Infections
RNA, Ribosomal, 16S
Sensitivity and Specificity
Uracil-DNA Glycosidase
วันที่เผยแพร่: 2018
บทคัดย่อ: Pseudomonas aeruginosa (P. aeruginosa) is an important opportunistic pathogen that causes serious infections in humans, including keratitis in contact lens wearers. Therefore, establishing a rapid, specific and sensitive method for the identification of P. aeruginosa is imperative. In the present study, the uracil-DNA-glycosylase-supplemented loop-mediated isothermal amplification combined with nanogold labeled hybridization probe (UDG-LAMP-AuNP) was developed for the detection of P. aeruginosa. UDG-LAMP was performed to prevent carry over contamination and the LAMP reactions can be readily observed using the nanogold probe. A set of 4 primers and a hybridization probe were designed based on the ecfX gene. The UDG-LAMP reactions were performed at 65°C for 60 min using the ratio of 40% deoxyuridine triphosphate to 60% deoxythymidine triphosphate. The detection of UDG-LAMP products using the nanogold labeled hybridization probe, which appeared as a red-purple color, was examined at 65°C for 5 min with 40 mM MgSO4. The UDG-LAMP-AuNP demonstrated specificity to all tested isolates of P. aeruginosa without cross reaction to other bacteria. The sensitivity for the detection of pure culture was 1.6×103 colony-forming units (CFU) ml-1 or equivalent to 3 CFU per reaction while that of polymerase chain reaction was 30 CFU per reaction. The detection limit of spiked contact lenses was 1.1×103 CFU ml-1 or equivalent to 2 CFU per reaction. In conclusion, the UDG-LAMP-AuNP assay was rapid, simple, specific and was effective for the identification of P. aeruginosa in contaminated samples. © 2018 Kashyap.
URI: https://ir.swu.ac.th/jspui/handle/123456789/13588
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85043294357&doi=10.3892%2fmmr.2018.8557&partnerID=40&md5=ca22d8f4dbd8e9920b007c635d5d157d
ISSN: 17912997
Appears in Collections:Scopus 1983-2021

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