A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics

This work describes the development of a Point-of-Care (POC) Lab-on-a-Chip (LOC) instrument for diagnosis of SARS-CoV-2 by Reverse-Transcription Loop-mediated isothermal amplification (RT-LAMP). The hardware is based on a Raspberry Pi computer ($35), a video camera, an Arduino Nano microcontroller,...

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Autores principales: Torezin Mendonça, Geovani, Cassaboni Stracke, Mateus, de Oliveira Coelho, Bruna, Bruna Soligo Sanchuki, Heloisa, Klassen de Oliveira, Viviane, Klerynton Marchini, Fabricio, Lucíola Zanette, Dalila, Nóbrega Aoki, Mateus, Ribeiro Viana, Emilson, Blanes, Lucas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9121651/
https://www.ncbi.nlm.nih.gov/pubmed/35620142
http://dx.doi.org/10.1016/j.microc.2022.107600
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author Torezin Mendonça, Geovani
Cassaboni Stracke, Mateus
de Oliveira Coelho, Bruna
Bruna Soligo Sanchuki, Heloisa
Klassen de Oliveira, Viviane
Klerynton Marchini, Fabricio
Lucíola Zanette, Dalila
Nóbrega Aoki, Mateus
Ribeiro Viana, Emilson
Blanes, Lucas
author_facet Torezin Mendonça, Geovani
Cassaboni Stracke, Mateus
de Oliveira Coelho, Bruna
Bruna Soligo Sanchuki, Heloisa
Klassen de Oliveira, Viviane
Klerynton Marchini, Fabricio
Lucíola Zanette, Dalila
Nóbrega Aoki, Mateus
Ribeiro Viana, Emilson
Blanes, Lucas
author_sort Torezin Mendonça, Geovani
collection PubMed
description This work describes the development of a Point-of-Care (POC) Lab-on-a-Chip (LOC) instrument for diagnosis of SARS-CoV-2 by Reverse-Transcription Loop-mediated isothermal amplification (RT-LAMP). The hardware is based on a Raspberry Pi computer ($35), a video camera, an Arduino Nano microcontroller, a printed circuit board as a heater and a 3D printed housing. The chips were manufactured in polymethyl methacrylate (PMMA) using a CO2 laser cutting machine and sealed with a PCR optic plastic film. The chip temperature is precisely controlled by a proportional–integral–derivative (PID) algorithm. During the RT-LAMP amplifications the chip was maintained at ∼ (65.0 ± 0.1) °C for 25 minutes and 5 minutes cooling down, totaling a 30 minutes of reaction .The software interpretation occurs in less than a second. The chip design has four 25 µL chambers, two for clinical samples and two for positive and negative control-samples. The RT-LAMP master mix solution added in the chip chambers contains the pH indicator Phenol Red, that is pink (for pH ∼ 8.0) before amplification and becomes yellow (pH ∼ 6.0) if the genetic material is amplified. The RT-LAMP SARS-CoV-2 diagnostic was made by color image recognition using the OpenCV machine vision software library. The software was programmed to automatically distinguish the HSV color parameter distribution in each one of the four chip chambers. The instrument was successfully tested for SARS-CoV-2 diagnosis, in 22 clinic samples, 11 positives and 11 negatives, achieving an assertiveness of 86% when compared to the results obtained by RT-LAMP standard reactions performed in conventional PCR equipment.
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spelling pubmed-91216512022-05-20 A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics Torezin Mendonça, Geovani Cassaboni Stracke, Mateus de Oliveira Coelho, Bruna Bruna Soligo Sanchuki, Heloisa Klassen de Oliveira, Viviane Klerynton Marchini, Fabricio Lucíola Zanette, Dalila Nóbrega Aoki, Mateus Ribeiro Viana, Emilson Blanes, Lucas Microchem J Article This work describes the development of a Point-of-Care (POC) Lab-on-a-Chip (LOC) instrument for diagnosis of SARS-CoV-2 by Reverse-Transcription Loop-mediated isothermal amplification (RT-LAMP). The hardware is based on a Raspberry Pi computer ($35), a video camera, an Arduino Nano microcontroller, a printed circuit board as a heater and a 3D printed housing. The chips were manufactured in polymethyl methacrylate (PMMA) using a CO2 laser cutting machine and sealed with a PCR optic plastic film. The chip temperature is precisely controlled by a proportional–integral–derivative (PID) algorithm. During the RT-LAMP amplifications the chip was maintained at ∼ (65.0 ± 0.1) °C for 25 minutes and 5 minutes cooling down, totaling a 30 minutes of reaction .The software interpretation occurs in less than a second. The chip design has four 25 µL chambers, two for clinical samples and two for positive and negative control-samples. The RT-LAMP master mix solution added in the chip chambers contains the pH indicator Phenol Red, that is pink (for pH ∼ 8.0) before amplification and becomes yellow (pH ∼ 6.0) if the genetic material is amplified. The RT-LAMP SARS-CoV-2 diagnostic was made by color image recognition using the OpenCV machine vision software library. The software was programmed to automatically distinguish the HSV color parameter distribution in each one of the four chip chambers. The instrument was successfully tested for SARS-CoV-2 diagnosis, in 22 clinic samples, 11 positives and 11 negatives, achieving an assertiveness of 86% when compared to the results obtained by RT-LAMP standard reactions performed in conventional PCR equipment. Elsevier B.V. 2022-09 2022-05-20 /pmc/articles/PMC9121651/ /pubmed/35620142 http://dx.doi.org/10.1016/j.microc.2022.107600 Text en © 2022 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Torezin Mendonça, Geovani
Cassaboni Stracke, Mateus
de Oliveira Coelho, Bruna
Bruna Soligo Sanchuki, Heloisa
Klassen de Oliveira, Viviane
Klerynton Marchini, Fabricio
Lucíola Zanette, Dalila
Nóbrega Aoki, Mateus
Ribeiro Viana, Emilson
Blanes, Lucas
A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title_full A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title_fullStr A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title_full_unstemmed A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title_short A new RT-LAMP-on-a-Chip Instrument for SARS-CoV-2 diagnostics
title_sort new rt-lamp-on-a-chip instrument for sars-cov-2 diagnostics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9121651/
https://www.ncbi.nlm.nih.gov/pubmed/35620142
http://dx.doi.org/10.1016/j.microc.2022.107600
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