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Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria

The polymerase chain reaction (PCR) is a robust technique used to make multiple copies of a segment of DNA. However, the available PCR platforms require elaborate and time-consuming operations or costly instruments, hindering their application. Herein, we introduce a sandwiched glass–polydimethylsil...

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Autores principales: Gorgannezhad, Lena, Sreejith, Kamalalayam Rajan, Zhang, Jun, Kijanka, Gregor, Christie, Melody, Stratton, Helen, Nguyen, Nam-Trung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6952809/
https://www.ncbi.nlm.nih.gov/pubmed/31888270
http://dx.doi.org/10.3390/mi10120883
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author Gorgannezhad, Lena
Sreejith, Kamalalayam Rajan
Zhang, Jun
Kijanka, Gregor
Christie, Melody
Stratton, Helen
Nguyen, Nam-Trung
author_facet Gorgannezhad, Lena
Sreejith, Kamalalayam Rajan
Zhang, Jun
Kijanka, Gregor
Christie, Melody
Stratton, Helen
Nguyen, Nam-Trung
author_sort Gorgannezhad, Lena
collection PubMed
description The polymerase chain reaction (PCR) is a robust technique used to make multiple copies of a segment of DNA. However, the available PCR platforms require elaborate and time-consuming operations or costly instruments, hindering their application. Herein, we introduce a sandwiched glass–polydimethylsiloxane (PDMS)–glass microchip containing an array of reactors for the real-time PCR-based detection of multiple waterborne bacteria. The PCR solution was loaded into the array of reactors in a single step utilising capillary filling, eliminating the need for pumps, valves, and liquid handling instruments. Issues of generating and trapping bubbles during the loading chip step were addressed by creating smooth internal reactor surfaces. Triton X-100 was used to enhance PCR compatibility in the chip by minimising the nonspecific adsorption of enzymes. A custom-made real-time PCR instrument was also fabricated to provide thermal cycling to the array chip. The microfluidic device was successfully demonstrated for microbial faecal source tracking (MST) in water.
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spelling pubmed-69528092020-01-23 Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria Gorgannezhad, Lena Sreejith, Kamalalayam Rajan Zhang, Jun Kijanka, Gregor Christie, Melody Stratton, Helen Nguyen, Nam-Trung Micromachines (Basel) Article The polymerase chain reaction (PCR) is a robust technique used to make multiple copies of a segment of DNA. However, the available PCR platforms require elaborate and time-consuming operations or costly instruments, hindering their application. Herein, we introduce a sandwiched glass–polydimethylsiloxane (PDMS)–glass microchip containing an array of reactors for the real-time PCR-based detection of multiple waterborne bacteria. The PCR solution was loaded into the array of reactors in a single step utilising capillary filling, eliminating the need for pumps, valves, and liquid handling instruments. Issues of generating and trapping bubbles during the loading chip step were addressed by creating smooth internal reactor surfaces. Triton X-100 was used to enhance PCR compatibility in the chip by minimising the nonspecific adsorption of enzymes. A custom-made real-time PCR instrument was also fabricated to provide thermal cycling to the array chip. The microfluidic device was successfully demonstrated for microbial faecal source tracking (MST) in water. MDPI 2019-12-16 /pmc/articles/PMC6952809/ /pubmed/31888270 http://dx.doi.org/10.3390/mi10120883 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gorgannezhad, Lena
Sreejith, Kamalalayam Rajan
Zhang, Jun
Kijanka, Gregor
Christie, Melody
Stratton, Helen
Nguyen, Nam-Trung
Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title_full Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title_fullStr Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title_full_unstemmed Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title_short Microfluidic Array Chip for Parallel Detection of Waterborne Bacteria
title_sort microfluidic array chip for parallel detection of waterborne bacteria
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6952809/
https://www.ncbi.nlm.nih.gov/pubmed/31888270
http://dx.doi.org/10.3390/mi10120883
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