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A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform

Objective: A Tag Array chip was used to detect plasmids of different template concentration, and then analyzed for sensitivity and specificity. Drug resistance genes from tuberculosis clinical specimens were detected, giving comparative phenotypic resistance results to explore the feasibility and va...

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Autores principales: Lyu, Jingtong, Wu, Wenjie, Cheng, Peng, Liu, Xun, Luo, Fei, Zhang, Zehua, Tang, Kanglai, Xu, Jianzhong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6143819/
https://www.ncbi.nlm.nih.gov/pubmed/30258420
http://dx.doi.org/10.3389/fmicb.2018.02106
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author Lyu, Jingtong
Wu, Wenjie
Cheng, Peng
Liu, Xun
Luo, Fei
Zhang, Zehua
Tang, Kanglai
Xu, Jianzhong
author_facet Lyu, Jingtong
Wu, Wenjie
Cheng, Peng
Liu, Xun
Luo, Fei
Zhang, Zehua
Tang, Kanglai
Xu, Jianzhong
author_sort Lyu, Jingtong
collection PubMed
description Objective: A Tag Array chip was used to detect plasmids of different template concentration, and then analyzed for sensitivity and specificity. Drug resistance genes from tuberculosis clinical specimens were detected, giving comparative phenotypic resistance results to explore the feasibility and value of clinical applications. Methods: Twenty-four strains of Mycobacterium Tuberculosis (MTB) having sequence differences in extracted plasmids of mutant strains. The plasmid was diluted into different concentrations, and then was performed to analyze the sensitivity and specificity of the chip system. A total of 427 clinical specimens (including spinal tuberculosis and pulmonary tuberculosis) were collected from patients who came from seven hospitals. Design, optimization and preparation of the chip detection system, sequencing and phenotypic drug susceptibility results were analyzed to evaluate the sensitivity and specificity of the gene chip. Results: In the template, concentrations of 1 × 10(3) copies/μL and above were consistent with sequencing results in the mutant. The sensitivity and specificity in spine Tuberculosis specimen of rifampicin (RFP) were 94.40 and 92.86%; isoniazide (INH) were 92.37 and 87.50%; ethambutol (EMB) were 61.36 and 89.29%; fluoroquinolones (FQS) were 79.41 and 92.86%; streptomycin (SM) were 90.18 and 89.29%; second line drugs (SLD) were 77.61 and 83.93%. In Pulmonary Tuberculosis specimen, the sensitivity and specificity respectively were RFP: 92.74%; 93.75%; INH: 91.26%; 87.50%; EMB: 54.17%; 89.58%; FQS: 84.87%; 93.75%; SM: 86.73%; 85.42%; SLD: 80.9%; 91.67%. The RFP, INH, FQs and SM resistance genes was highly sensitive and specific: however, for detection of amikacin (AMK), capreomycin (CPM), kanamycin (KM), specificity was higher, but sensitivity was lower. Sensitivity for the detection of a mutation in the eis promoter region could be improved. Conclusion: Tag Array chip can achieve rapid, accurate detection of tuberculosis resistance, which has important clinical significance and feasibility.
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spelling pubmed-61438192018-09-26 A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform Lyu, Jingtong Wu, Wenjie Cheng, Peng Liu, Xun Luo, Fei Zhang, Zehua Tang, Kanglai Xu, Jianzhong Front Microbiol Microbiology Objective: A Tag Array chip was used to detect plasmids of different template concentration, and then analyzed for sensitivity and specificity. Drug resistance genes from tuberculosis clinical specimens were detected, giving comparative phenotypic resistance results to explore the feasibility and value of clinical applications. Methods: Twenty-four strains of Mycobacterium Tuberculosis (MTB) having sequence differences in extracted plasmids of mutant strains. The plasmid was diluted into different concentrations, and then was performed to analyze the sensitivity and specificity of the chip system. A total of 427 clinical specimens (including spinal tuberculosis and pulmonary tuberculosis) were collected from patients who came from seven hospitals. Design, optimization and preparation of the chip detection system, sequencing and phenotypic drug susceptibility results were analyzed to evaluate the sensitivity and specificity of the gene chip. Results: In the template, concentrations of 1 × 10(3) copies/μL and above were consistent with sequencing results in the mutant. The sensitivity and specificity in spine Tuberculosis specimen of rifampicin (RFP) were 94.40 and 92.86%; isoniazide (INH) were 92.37 and 87.50%; ethambutol (EMB) were 61.36 and 89.29%; fluoroquinolones (FQS) were 79.41 and 92.86%; streptomycin (SM) were 90.18 and 89.29%; second line drugs (SLD) were 77.61 and 83.93%. In Pulmonary Tuberculosis specimen, the sensitivity and specificity respectively were RFP: 92.74%; 93.75%; INH: 91.26%; 87.50%; EMB: 54.17%; 89.58%; FQS: 84.87%; 93.75%; SM: 86.73%; 85.42%; SLD: 80.9%; 91.67%. The RFP, INH, FQs and SM resistance genes was highly sensitive and specific: however, for detection of amikacin (AMK), capreomycin (CPM), kanamycin (KM), specificity was higher, but sensitivity was lower. Sensitivity for the detection of a mutation in the eis promoter region could be improved. Conclusion: Tag Array chip can achieve rapid, accurate detection of tuberculosis resistance, which has important clinical significance and feasibility. Frontiers Media S.A. 2018-09-07 /pmc/articles/PMC6143819/ /pubmed/30258420 http://dx.doi.org/10.3389/fmicb.2018.02106 Text en Copyright © 2018 Lyu, Wu, Cheng, Liu, Luo, Zhang, Tang and Xu. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Lyu, Jingtong
Wu, Wenjie
Cheng, Peng
Liu, Xun
Luo, Fei
Zhang, Zehua
Tang, Kanglai
Xu, Jianzhong
A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title_full A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title_fullStr A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title_full_unstemmed A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title_short A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform
title_sort chip for detecting tuberculosis drug resistance based on polymerase chain reaction (pcr)-magnetic bead molecule platform
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6143819/
https://www.ncbi.nlm.nih.gov/pubmed/30258420
http://dx.doi.org/10.3389/fmicb.2018.02106
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