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Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA

Loop-mediated isothermal amplification (LAMP) is a promising diagnostic tool for genetic amplification, which is known for its rapid process, simple operation, high amplification efficiency, and excellent sensitivity. However, most of the existing heating methods are external for completion of molec...

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Autores principales: Zhu, Huan-Huan, Li, Yuan, Wu, Li-Xia, Wang, Ke-Sheng, Zhang, Yue, Fan, Qiang-Yuan, Ming, Zun-Zhen, Chen, Wei-Qin, Liu, Wei-Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Vienna 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065241/
https://www.ncbi.nlm.nih.gov/pubmed/35507110
http://dx.doi.org/10.1007/s00604-022-05283-9
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author Zhu, Huan-Huan
Li, Yuan
Wu, Li-Xia
Wang, Ke-Sheng
Zhang, Yue
Fan, Qiang-Yuan
Ming, Zun-Zhen
Chen, Wei-Qin
Liu, Wei-Wei
author_facet Zhu, Huan-Huan
Li, Yuan
Wu, Li-Xia
Wang, Ke-Sheng
Zhang, Yue
Fan, Qiang-Yuan
Ming, Zun-Zhen
Chen, Wei-Qin
Liu, Wei-Wei
author_sort Zhu, Huan-Huan
collection PubMed
description Loop-mediated isothermal amplification (LAMP) is a promising diagnostic tool for genetic amplification, which is known for its rapid process, simple operation, high amplification efficiency, and excellent sensitivity. However, most of the existing heating methods are external for completion of molecular amplification with possibility of contamination of specimens. The present research provided an internal heating method for LAMP using magnetic nanoparticles (MNPs), which is called nano-LAMP. Near-infrared light with an excitation wavelength of 808 nm was employed as the heating source; hydroxy naphthol blue (HNB) was used as an indicator to conduct methodological research. We demonstrate that the best temperature was controlled at a working power of 2 W and 4.8 µg/µL concentration of nanoparticles. The lowest limit for the detection of HPV by the nano-LAMP method is 10(2) copies/mL, which was confirmed by a gel electrophoresis assay. In the feasibility investigation of validated clinical samples, all 10 positive HPV-6 specimens amplified by nano-LAMP were consistent with conventional LAMP methods. Therefore, the nano-LAMP detection method using internal heating of MNPs may bring a new vision to the exploration of thermostatic detection in the future. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-022-05283-9.
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spelling pubmed-90652412022-05-04 Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA Zhu, Huan-Huan Li, Yuan Wu, Li-Xia Wang, Ke-Sheng Zhang, Yue Fan, Qiang-Yuan Ming, Zun-Zhen Chen, Wei-Qin Liu, Wei-Wei Mikrochim Acta Original Paper Loop-mediated isothermal amplification (LAMP) is a promising diagnostic tool for genetic amplification, which is known for its rapid process, simple operation, high amplification efficiency, and excellent sensitivity. However, most of the existing heating methods are external for completion of molecular amplification with possibility of contamination of specimens. The present research provided an internal heating method for LAMP using magnetic nanoparticles (MNPs), which is called nano-LAMP. Near-infrared light with an excitation wavelength of 808 nm was employed as the heating source; hydroxy naphthol blue (HNB) was used as an indicator to conduct methodological research. We demonstrate that the best temperature was controlled at a working power of 2 W and 4.8 µg/µL concentration of nanoparticles. The lowest limit for the detection of HPV by the nano-LAMP method is 10(2) copies/mL, which was confirmed by a gel electrophoresis assay. In the feasibility investigation of validated clinical samples, all 10 positive HPV-6 specimens amplified by nano-LAMP were consistent with conventional LAMP methods. Therefore, the nano-LAMP detection method using internal heating of MNPs may bring a new vision to the exploration of thermostatic detection in the future. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-022-05283-9. Springer Vienna 2022-05-04 2022 /pmc/articles/PMC9065241/ /pubmed/35507110 http://dx.doi.org/10.1007/s00604-022-05283-9 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature 2022 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Paper
Zhu, Huan-Huan
Li, Yuan
Wu, Li-Xia
Wang, Ke-Sheng
Zhang, Yue
Fan, Qiang-Yuan
Ming, Zun-Zhen
Chen, Wei-Qin
Liu, Wei-Wei
Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title_full Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title_fullStr Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title_full_unstemmed Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title_short Internal heating method of loop-mediated isothermal amplification for detection of HPV-6 DNA
title_sort internal heating method of loop-mediated isothermal amplification for detection of hpv-6 dna
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065241/
https://www.ncbi.nlm.nih.gov/pubmed/35507110
http://dx.doi.org/10.1007/s00604-022-05283-9
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