Publication:
Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples

3
0

Issued Date

2013

Resource Type

Article

File Type

application/pdf

ISSN

23146133

DOI

10.1155/2013/926230

Other identifier(s)

2-s2.0-84875754885

Rights Holder(s)

Scopus

Bibliographic Citation

BioMed Research International. Vol 2013, No. (2013), p.-

Suggested Citation

Kaewphinit T., Arunrut N., Kiatpathomchai W., Santiwatanakul S., Jaratsing P., Chansiri K. Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples. BioMed Research International. Vol 2013, No. (2013), p.-. doi:10.1155/2013/926230 Retrieved from: https://hdl.handle.net/20.500.14740/6680

Title

Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples

Author(s)

Kaewphinit T.
 Arunrut N.
 Kiatpathomchai W.
 Santiwatanakul S.
 Jaratsing P.
 Chansiri K.

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.

Subject(s)

Bacterial DNA
 Fluorescein isothiocyanate
 Genomic DNA
 Oligonucleotide
 Bacterial DNA
 Primer DNA
 Article
 Bacterial gene
 Bacterium culture
 Bioassay
 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
 Nonhuman
 Nucleotide sequence
 Sensitivity and specificity
 Chromatography
 Genetics
 Human
 Isolation and purification
 Microbiology
 Nucleic acid amplification
 Nucleic acid hybridization
 Pathology
 Species difference
 Tuberculosis
 Bacteria (microorganisms)
 Mycobacterium tuberculosis
 Chromatography
 DNA Primers
 DNA, Bacterial
 Humans
 Mycobacterium tuberculosis
 Nucleic Acid Amplification Techniques
 Nucleic Acid Hybridization
 Species Specificity
 Tuberculosis

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URI

https://hdl.handle.net/20.500.14740/6680

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Scopus: Year 1983-2021

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