MGIT-seq for the Identification of Nontuberculous Mycobacteria and Drug Resistance: a Prospective Study

Fukushima, Kiyoharu; Matsumoto, Yuki; Saito, Haruko; Motooka, Daisuke; Komukai, Sho; Fukui, Emiko; Yamuchi, June; Nitta, Tadayoshi; Niitsu, Takayuki; Abe, Yuko; Nabeshima, Hiroshi; Nagahama, Yasuharu; Nii, Takuro; Tsujino, Kazuyuki; Miki, Keisuke; Kitada, Shuichi; Kumanogoh, Atsushi; Akira, Shizuo; Nakamura, Shota; Kida, Hiroshi

doi:10.1128/jcm.01626-22

J Clin Microbiol

2023

DOI: 10.1128/jcm.01626-22

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MGIT-seq for the Identification of Nontuberculous Mycobacteria and Drug Resistance: a Prospective Study

Kiyoharu Fukushima

Yuki Matsumoto

Haruko Saito

et al.

Abstract: Because nontuberculous mycobacterial pulmonary disease is a considerable health burden, a simple and clinically applicable analytical protocol enabling the identification of subspecies and drug-resistant disease is required to determine the treatment strategy. We aimed to develop a simplified workflow consisting only of direct sequencing of mycobacterial growth indicator tube cultures (MGIT-seq).

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Cited by 4 publications

References 61 publications

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Integrating Paper‐Based Microfluidics and Lateral Flow Strip into Nucleic Acid Amplification Device toward Rapid, Low‐Cost, and Visual Diagnosis of Multiple Mycobacteria

Wu,

Shen,

Zhao

et al. 2024

Small Methods

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Efficient diagnosis of mycobacterial infections can effectively manage and prevent the transmission of infectious diseases. Unfortunately, existing diagnostic strategies are challenged by long assay times, high costs, and highly specialized expertise to distinguish between pulmonary tuberculosis (PTB) and nontuberculous mycobacterial pulmonary diseases (NTM‐PDs). Herein, in this study, an optimized 3D paper‐based analytical device (µPAD) is incorporated with a closed lateral flow (LF) strip into a loop‐mediated isothermal amplification (LAMP) device (3D‐µPAD‐LF‐LAMP) for rapid, low‐cost, and visual detection of pathogenic mycobacteria. The platform's microfluidic feature enhanced the nucleic acid amplification, thereby reducing the costs and time as compared to boiling, easyMAG, and QIAGEN techniques. Moreover, the LF unit is specifically designed to minimize aerosol contamination for a user‐friendly and visual readout. 3D‐µPAD‐LF‐LAMP is optimized and assessed using standard strains, demonstrating a limit of detection (LOD) down to 10 fg reaction−1. In a cohort of 815 patients, 3D‐µPAD‐LF‐LAMP displays significantly better sensitivity, specificity, negative predictive value (NPV), positive predictive value (PPV), and diagnostic accuracy than conventional bacterial culture and Xpert techniques. Collectively, 3D‐µPAD‐LF‐LAMP demonstrates enhanced accessibility, efficiency, and practicality for the diagnosis of multiple pathogenic mycobacteria, which can be applied across diverse clinical settings, thereby ultimately improving public health outcomes.

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Integrating Paper‐Based Microfluidics and Lateral Flow Strip into Nucleic Acid Amplification Device toward Rapid, Low‐Cost, and Visual Diagnosis of Multiple Mycobacteria

Wu,

Shen,

Zhao

et al. 2024

Small Methods

View full text Add to dashboard Cite

show abstract

Current Trends in Identification and Susceptibility Testing of Mycobacteria

Thwe,

Rajagopalan,

Orner

2024

Clinics in Laboratory Medicine

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Evaluation of the GenoType NTM-DR line probe assay for nontuberculous mycobacteria using whole genome sequences as reference standard

Epperson,

Davidson,

Kammlade

et al. 2024

Diagnostic Microbiology and Infectious Disease

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MGIT-seq for the Identification of Nontuberculous Mycobacteria and Drug Resistance: a Prospective Study

Cited by 4 publications

References 61 publications

Integrating Paper‐Based Microfluidics and Lateral Flow Strip into Nucleic Acid Amplification Device toward Rapid, Low‐Cost, and Visual Diagnosis of Multiple Mycobacteria

Integrating Paper‐Based Microfluidics and Lateral Flow Strip into Nucleic Acid Amplification Device toward Rapid, Low‐Cost, and Visual Diagnosis of Multiple Mycobacteria

Current Trends in Identification and Susceptibility Testing of Mycobacteria

Evaluation of the GenoType NTM-DR line probe assay for nontuberculous mycobacteria using whole genome sequences as reference standard

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