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,...
Autores principales: | , , , , , , , , , |
---|---|
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 |
_version_ | 1784711195920433152 |
---|---|
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. |
format | Online Article Text |
id | pubmed-9121651 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT torezinmendoncageovani anewrtlamponachipinstrumentforsarscov2diagnostics AT cassabonistrackemateus anewrtlamponachipinstrumentforsarscov2diagnostics AT deoliveiracoelhobruna anewrtlamponachipinstrumentforsarscov2diagnostics AT brunasoligosanchukiheloisa anewrtlamponachipinstrumentforsarscov2diagnostics AT klassendeoliveiraviviane anewrtlamponachipinstrumentforsarscov2diagnostics AT kleryntonmarchinifabricio anewrtlamponachipinstrumentforsarscov2diagnostics AT luciolazanettedalila anewrtlamponachipinstrumentforsarscov2diagnostics AT nobregaaokimateus anewrtlamponachipinstrumentforsarscov2diagnostics AT ribeirovianaemilson anewrtlamponachipinstrumentforsarscov2diagnostics AT blaneslucas anewrtlamponachipinstrumentforsarscov2diagnostics AT torezinmendoncageovani newrtlamponachipinstrumentforsarscov2diagnostics AT cassabonistrackemateus newrtlamponachipinstrumentforsarscov2diagnostics AT deoliveiracoelhobruna newrtlamponachipinstrumentforsarscov2diagnostics AT brunasoligosanchukiheloisa newrtlamponachipinstrumentforsarscov2diagnostics AT klassendeoliveiraviviane newrtlamponachipinstrumentforsarscov2diagnostics AT kleryntonmarchinifabricio newrtlamponachipinstrumentforsarscov2diagnostics AT luciolazanettedalila newrtlamponachipinstrumentforsarscov2diagnostics AT nobregaaokimateus newrtlamponachipinstrumentforsarscov2diagnostics AT ribeirovianaemilson newrtlamponachipinstrumentforsarscov2diagnostics AT blaneslucas newrtlamponachipinstrumentforsarscov2diagnostics |