TY - JOUR
T1 - Rapid and simple detection of isoniazid-resistant mycobacterium tuberculosis utilizing a dna chromatography-based technique
AU - Kodera, Takuya
AU - Yamaguchi, Tomoyuki
AU - Fukushima, Yukari
AU - Kobayashi, Kumi
AU - Takarada, Yutaka
AU - Chizimu, Joseph Yamweka
AU - Nakajima, Chie
AU - Solo, Eddie Samuneti
AU - Lungu, Patrick Saili
AU - Kawase, Mitsuo
AU - Suzuki, Yasuhiko
N1 - Funding Information:
The authors would like to thank Dr. Aki Tamaru for her support to provide positive control DNA. The authors also appreciate Japan BCG Laboratory for providing a bacterial strain. This work was supported in part by a grant from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan, for the Joint Research Program of the Research Center for Zoonosis Control, Hokkaido University to YS, and in part by Japan Agency for Medical Research and Development (AMED) under Grant Number JP20jk0210005, JP20jm0110021, and JP20wm0125008 to YS.
Publisher Copyright:
© 2021, National Institute of Health. All rights reserved.
PY - 2021
Y1 - 2021
N2 - Despite the availability of anti-tuberculosis drugs, the treatment of tuberculosis has been complicated by drug-resistant tuberculosis. The early detection of drug resistance makes early treatment possible. However, the available tools are mainly for rifampicin resistance detection, and the existing isoniazid resistance detection method is expensive, highly technical, and complicated, making it unsustainable for use in developing nations. This study aimed to develop a simple, rapid, and low-cost diagnostic kit for isoniazid-resistant tuberculosis using the single-stranded tag hybridization method to target an isoniazid resistance-conferring mutation. Specificity and sensitivity were assessed using DNA extracted from 49 isoniazid-resistant and 41 isoniazid-susceptible Mycobacterium tuberculosis clinical isolates cultured in mycobacterial growth indicator tubes. Positive signals were observed on mutant and wild-type lines with 100% sensitivity and specificity compared with Sanger sequencing results. In contrast, no positive signal was observed for non-tuberculosis mycobacteria. The detection limit of this method was 103 CFU or less. The STH-PAS system for isoniazid-resistant M. tuberculosis detection developed in this study offers a better alternative to conventional phenotypic isoniazid resistance determination, which will be of both clinical and epidemiological significance in resource-limited nations.
AB - Despite the availability of anti-tuberculosis drugs, the treatment of tuberculosis has been complicated by drug-resistant tuberculosis. The early detection of drug resistance makes early treatment possible. However, the available tools are mainly for rifampicin resistance detection, and the existing isoniazid resistance detection method is expensive, highly technical, and complicated, making it unsustainable for use in developing nations. This study aimed to develop a simple, rapid, and low-cost diagnostic kit for isoniazid-resistant tuberculosis using the single-stranded tag hybridization method to target an isoniazid resistance-conferring mutation. Specificity and sensitivity were assessed using DNA extracted from 49 isoniazid-resistant and 41 isoniazid-susceptible Mycobacterium tuberculosis clinical isolates cultured in mycobacterial growth indicator tubes. Positive signals were observed on mutant and wild-type lines with 100% sensitivity and specificity compared with Sanger sequencing results. In contrast, no positive signal was observed for non-tuberculosis mycobacteria. The detection limit of this method was 103 CFU or less. The STH-PAS system for isoniazid-resistant M. tuberculosis detection developed in this study offers a better alternative to conventional phenotypic isoniazid resistance determination, which will be of both clinical and epidemiological significance in resource-limited nations.
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U2 - 10.7883/yoken.JJID.2020.754
DO - 10.7883/yoken.JJID.2020.754
M3 - Article
C2 - 33132303
AN - SCOPUS:85107088211
SN - 1344-6304
VL - 74
SP - 214
EP - 219
JO - Japanese medical journal
JF - Japanese medical journal
IS - 3
ER -
