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Title: Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples
Authors: Kaewphinit T.
Arunrut N.
Kiatpathomchai W.
Santiwatanakul S.
Jaratsing P.
Chansiri K.
Keywords: bacterial DNA
fluorescein isothiocyanate
genomic DNA
bacterial DNA
primer DNA
bacterial gene
bacterium culture
controlled study
cost effectiveness analysis
DNA extraction
DNA hybridization
lateral flow dipstick
loop mediated isothermal amplification
Mycobacterium avium
Mycobacterium fortuitum
Mycobacterium gordonae
Mycobacterium intracellulare
Mycobacterium kansasii
Mycobacterium tuberculosis
nucleotide sequence
sensitivity and specificity
isolation and purification
nucleic acid amplification
nucleic acid hybridization
species difference
Bacteria (microorganisms)
Mycobacterium tuberculosis
DNA Primers
DNA, Bacterial
Mycobacterium tuberculosis
Nucleic Acid Amplification Techniques
Nucleic Acid Hybridization
Species Specificity
Issue Date: 2013
Abstract: Tuberculosis (TB) is a communicable disease caused by the bacterium Mycobacterium tuberculosis (MTB) and is a persistent problem in the developing countries. Loop-mediated isothermal amplification (LAMP) allows DNA to be amplified rapidly at a constant temperature. Here, a LAMP method was combined with a chromatographic lateral-flow dipstick (LFD) to detect IS6110 gene of M. tuberculosis specifically and rapidly. The reaction was optimized at 63°C for 60 min, and the amplified DNA hybridized to an FITC-labeled oligonucleotide probe for 5 min was detected at the LFD test line 5 min after application. Excluding the step of DNA extraction, the test results could be generated approximately within 1 h. In addition to the advantage of short assay time, this technique could avoid the contact of carcinogenic ethidium bromide due to the exclusion of the electrophoresis analysis step. Furthermore, the data indicated that LAMP-LFD could detect M. tuberculosis genomic DNA as little as 5 pg. The technique showed a significant specificity since no cross-hybridization to M. intracellulare (MIC), M. fortuitum (MFT), M. avium (MAV), M. kansasii (MKS), and M. gordonae (MGD) genomic DNAs was observed. In the clinical unknown samples test, the sensitivity of LAMP-LFD was 98.92% and the specificity was 100% compared to those of the standard culture assay. Based on its sensitivity, specificity, rapidity, low cost, and convenience, LAMP-LFD could be applicable for use in both laboratories and epidemiological surveys of MTB. © 2013 Thongchai Kaewphinit et al.
ISSN: 23146133
Appears in Collections:SCOPUS 1983-2021

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