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Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis

Proficiency testing based on quality control materials is an important component of the quality assurance system for detection methods. However, in the detection of infectious diseases, it is a challenge to use quality control materials derived from clinical samples or pathogens owing to their infec...

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Autores principales: Guan, Zehao, Han, Xuefei, Huang, Weigang, Wang, Xueliang, Wang, Hualiang, Fan, Yun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10063913/
https://www.ncbi.nlm.nih.gov/pubmed/37009507
http://dx.doi.org/10.3389/fcimb.2023.1128337
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author Guan, Zehao
Han, Xuefei
Huang, Weigang
Wang, Xueliang
Wang, Hualiang
Fan, Yun
author_facet Guan, Zehao
Han, Xuefei
Huang, Weigang
Wang, Xueliang
Wang, Hualiang
Fan, Yun
author_sort Guan, Zehao
collection PubMed
description Proficiency testing based on quality control materials is an important component of the quality assurance system for detection methods. However, in the detection of infectious diseases, it is a challenge to use quality control materials derived from clinical samples or pathogens owing to their infectious nature. The Xpert MTB/RIF assay, endorsed by the World Health Organization, is one of the most widely implemented assays in the detection of Mycobacterium tuberculosis along with rifampicin resistance and its heterogeneity. Clinical isolates are typically used as quality controls for this assay, leading to concerns about biosafety, constrained target sequence polymorphisms, and time-consuming preparation. In this study, a heterogeneous quality control library for the Xpert MTB/RIF assay was constructed based on DNA synthesis and site-directed mutation, which provides sufficient rifampicin resistance polymorphisms, enabling monitoring all five probes of Xpert MTB/RIF and its combinations. Escherichia coli and Bacillus subtilis were used as heterogeneous hosts rather than the pathogen itself to eliminate biosafety risks; thus, preparation does not require a biosafety level III laboratory and the production time is reduced from a few months to a few days. The panel was stable for more than 15 months stored at 4°C and could be distributed at room temperature. All 11 laboratories in Shanghai participating in a pilot survey identified the specimens with corresponding probe patterns, and discordant results highlighted inappropriate operations in the process. Collectively, we show, for the first time, that this library, based on heterogeneous hosts, is an appropriate alternative for M. tuberculosis detection.
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spelling pubmed-100639132023-04-01 Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis Guan, Zehao Han, Xuefei Huang, Weigang Wang, Xueliang Wang, Hualiang Fan, Yun Front Cell Infect Microbiol Cellular and Infection Microbiology Proficiency testing based on quality control materials is an important component of the quality assurance system for detection methods. However, in the detection of infectious diseases, it is a challenge to use quality control materials derived from clinical samples or pathogens owing to their infectious nature. The Xpert MTB/RIF assay, endorsed by the World Health Organization, is one of the most widely implemented assays in the detection of Mycobacterium tuberculosis along with rifampicin resistance and its heterogeneity. Clinical isolates are typically used as quality controls for this assay, leading to concerns about biosafety, constrained target sequence polymorphisms, and time-consuming preparation. In this study, a heterogeneous quality control library for the Xpert MTB/RIF assay was constructed based on DNA synthesis and site-directed mutation, which provides sufficient rifampicin resistance polymorphisms, enabling monitoring all five probes of Xpert MTB/RIF and its combinations. Escherichia coli and Bacillus subtilis were used as heterogeneous hosts rather than the pathogen itself to eliminate biosafety risks; thus, preparation does not require a biosafety level III laboratory and the production time is reduced from a few months to a few days. The panel was stable for more than 15 months stored at 4°C and could be distributed at room temperature. All 11 laboratories in Shanghai participating in a pilot survey identified the specimens with corresponding probe patterns, and discordant results highlighted inappropriate operations in the process. Collectively, we show, for the first time, that this library, based on heterogeneous hosts, is an appropriate alternative for M. tuberculosis detection. Frontiers Media S.A. 2023-03-17 /pmc/articles/PMC10063913/ /pubmed/37009507 http://dx.doi.org/10.3389/fcimb.2023.1128337 Text en Copyright © 2023 Guan, Han, Huang, Wang, Wang and Fan https://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 Cellular and Infection Microbiology
Guan, Zehao
Han, Xuefei
Huang, Weigang
Wang, Xueliang
Wang, Hualiang
Fan, Yun
Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title_full Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title_fullStr Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title_full_unstemmed Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title_short Construction and application of a heterogeneous quality control library for the Xpert MTB/RIF assay in tuberculosis diagnosis
title_sort construction and application of a heterogeneous quality control library for the xpert mtb/rif assay in tuberculosis diagnosis
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10063913/
https://www.ncbi.nlm.nih.gov/pubmed/37009507
http://dx.doi.org/10.3389/fcimb.2023.1128337
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