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Ultrasensitive detection of Mycobacterium tuberculosis by a rapid and specific probe-triggered one-step, simultaneous DNA hybridization and isothermal amplification combined with a lateral flow dipstick.
Jaroenram, Wansadaj; Kampeera, Jantana; Arunrut, Narong; Sirithammajak, Sarawut; Jaitrong, Sarinya; Boonnak, Kobporn; Khumwan, Pakapreud; Prammananan, Therdsak; Chaiprasert, Angkana; Kiatpathomchai, Wansika.
Afiliação
  • Jaroenram W; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Kampeera J; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Arunrut N; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Sirithammajak S; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Jaitrong S; Tuberculosis Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Boonnak K; Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand.
  • Khumwan P; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Prammananan T; Tuberculosis Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
  • Chaiprasert A; Drug Resistant Tuberculosis Fund, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.
  • Kiatpathomchai W; Bioengineering and Sensing Technology Research Team, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, 12120, Thailand. wansika@biotec.or.th.
Sci Rep ; 10(1): 16976, 2020 10 12.
Article em En | MEDLINE | ID: mdl-33046776
Mycobacterium tuberculosis (Mtb) is an insidious scourge that has afflicted millions of people worldwide. Although there are many rapid methods to detect it based on loop-mediated isothermal amplification (LAMP) and a lateral flow dipstick (LFD), this study made further improvements using a new set of primers to enhance LAMP performance and a novel DNA probe system to simplify detection and increase specificity. The new probe system eliminates the post-LAMP hybridization step typically required for LFD assays by allowing co-hybridization and amplification of target DNA in one reaction while preventing self-polymerization that could lead to false-positive results. The improved assay was named Probe-Triggered, One-Step, Simultaneous DNA Hybridization and LAMP Integrated with LFD (SH-LAMP-LFD). SH-LAMP-LFD was simpler to perform and more sensitive than previously reported LAMP-LFD and PCR methods by 100 and 1000 times, respectively. It could detect a single cell of Mtb. The absence of cross-reactivity with 23 non-TB bacteria, and accurate test results with all 104 blind clinical samples have highlighted its accuracy. Its robustness and portability make SH-LAMP-LFD suitable for users in both low and high resource settings.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Tuberculose / DNA Bacteriano / Técnicas de Amplificação de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Mycobacterium tuberculosis / Hibridização de Ácido Nucleico Tipo de estudo: Clinical_trials / Diagnostic_studies Limite: Humans Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Tuberculose / DNA Bacteriano / Técnicas de Amplificação de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Mycobacterium tuberculosis / Hibridização de Ácido Nucleico Tipo de estudo: Clinical_trials / Diagnostic_studies Limite: Humans Idioma: En Ano de publicação: 2020 Tipo de documento: Article