Development of RNA/DNA automated extraction and purification device for infectious disease diagnosis.

COVID-19 Column purification method DNA PCR RNA

Journal

Practical laboratory medicine
ISSN: 2352-5517
Titre abrégé: Pract Lab Med
Pays: Netherlands
ID NLM: 101690848

Informations de publication

Date de publication:
Nov 2023
Historique:
received: 09 08 2023
revised: 02 09 2023
accepted: 03 09 2023
medline: 11 9 2023
pubmed: 11 9 2023
entrez: 11 9 2023
Statut: epublish

Résumé

Genetic tests using RNA/DNA are the most accurate for diagnosing infectious diseases and assessing disease susceptibility, including COVID-19. However, manual specimen handling and the risk of secondary infections by medical staff highlight the need for automated equipment. Automation methods, such as bead purification, have limitations with high-viscosity specimens, while column purification requires complex equipment. This study aimed to develop an automated device using the column purification method for safe and reliable infectious disease diagnosis. We compared the yield and purification of three nucleic acid extraction methods (centrifugation, pressurization, and depressurization) and examined the adaptation of the extraction methods to automated device. Furthermore, we examined the feasibility of extracting SARS-CoV-2 RNA from COVID-19 patients and using qPCR analysis to determine whether the extraction method could be used as a clinical analyzer. Results varied with different columns and reagents, but pressurization method was selected for the automated device's RNA/DNA extraction. Using an automated device equipped with a pressurization method, RNA extracted from pharyngeal fluids from COVID-19 patients who had already been diagnosed with SARS-CoV-2 by qRT-PCR again tested positive. These findings demonstrate the device's effectiveness for nucleic acid extraction and virus-targeted diagnostics. Moreover, it holds potential for genetic testing in fields like food and environmental measurements. The automated device addresses specimen handling challenges and provides a reliable tool for infectious disease diagnosis.

Identifiants

pubmed: 37693633
doi: 10.1016/j.plabm.2023.e00335
pii: S2352-5517(23)00029-X
pmc: PMC10492190
doi:

Types de publication

Journal Article

Langues

eng

Pagination

e00335

Informations de copyright

© 2023 The Authors.

Déclaration de conflit d'intérêts

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Références

J Biomed Biotechnol. 2009;2009:574398
pubmed: 20011662
Sci Rep. 2021 Mar 18;11(1):6260
pubmed: 33737536
Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15149-54
pubmed: 11742071
Sci Rep. 2018 Apr 3;8(1):5467
pubmed: 29615736
Handb Clin Neurol. 2014;123:123-47
pubmed: 25015483
Biomed Microdevices. 2009 Apr;11(2):339-50
pubmed: 19034667
Appl Environ Microbiol. 2005 Sep;71(9):5544-50
pubmed: 16151147
Nat Protoc. 2008;3(5):877-82
pubmed: 18451795
Emerg Microbes Infect. 2020 Dec;9(1):1744-1747
pubmed: 32673522
J Clin Microbiol. 2020 May 26;58(6):
pubmed: 32245835
Clin Chem. 1997 Nov;43(11):2021-38
pubmed: 9365385
Am J Hum Genet. 2021 Jul 1;108(7):1231-1238
pubmed: 34089648
Saf Health Work. 2020 Sep;11(3):372-377
pubmed: 32837739

Auteurs

Kyosuke Mukae (K)

Research Institute for Clinical Oncology, Saitama Cancer Center, Saitama, 362-0806, Japan.

Osamu Takei (O)

Research Institute for Clinical Oncology, Saitama Cancer Center, Saitama, 362-0806, Japan.
PMT Corporation, Fukuoka, 811-2115, Japan.

Fumi Imai (F)

Department of Clinical Laboratory Medicine, Saitama Cancer Center, Saitama, 362-0806, Japan.

Takehiko Kamijo (T)

Research Institute for Clinical Oncology, Saitama Cancer Center, Saitama, 362-0806, Japan.

Classifications MeSH