Use of high-resolution fluorescence in situ hybridization for fast and robust detection of SARS-CoV-2 RNAs.

Humans SARS-CoV-2 / genetics In Situ Hybridization, Fluorescence / methods RNA, Viral / genetics COVID-19 / diagnosis HEK293 Cells Phosphoproteins / genetics Coronavirus Envelope Proteins / genetics RNA, Messenger / genetics Coronavirus Nucleocapsid Proteins / genetics
COVID-19 Fluorescence in situ hybridization Infection RNA virus SARS-CoV-2 mRNA

Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
08 Sep 2024
Historique:
received: 10 05 2024
accepted: 22 08 2024
medline: 9 9 2024
pubmed: 9 9 2024
entrez: 8 9 2024
Statut: epublish

Résumé

Early, rapid, and accurate diagnostic tests play critical roles not only in the identification/management of individuals infected by SARS-CoV-2, but also in fast and effective public health surveillance, containment, and response. Our aim has been to develop a fast and robust fluorescence in situ hybridization (FISH) detection method for detecting SARS-CoV-2 RNAs by using an HEK 293 T cell culture model. At various times after being transfected with SARS-CoV-2 E and N plasmids, HEK 293 T cells were fixed and then hybridized with ATTO-labeled short DNA probes (about 20 nt). At 4 h, 12 h, and 24 h after transfection, SARS-CoV-2 E and N mRNAs were clearly revealed as solid granular staining inside HEK 293 T cells at all time points. Hybridization time was also reduced to 1 h for faster detection, and the test was completed within 3 h with excellent results. In addition, we have successfully detected 3 mRNAs (E mRNA, N mRNA, and ORF1a (-) RNA) simultaneously inside the buccal cells of COVID-19 patients. Our high-resolution RNA FISH might significantly increase the accuracy and efficiency of SARS-CoV-2 detection, while significantly reducing test time. The method can be conducted on smears containing cells (e.g., from nasopharyngeal, oropharyngeal, or buccal swabs) or smears without cells (e.g., from sputum, saliva, or drinking water/wastewater) for detecting various types of RNA viruses and even DNA viruses at different timepoints of infection.

Identifiants

DOI: 10.1038/s41598-024-70980-9 PMID: 39245656
pubmed: 39245656
doi: 10.1038/s41598-024-70980-9
pii: 10.1038/s41598-024-70980-9
doi:

Substances chimiques

RNA, Viral 0
Phosphoproteins 0
Coronavirus Envelope Proteins 0
RNA, Messenger 0
nucleocapsid phosphoprotein, SARS-CoV-2 0
Coronavirus Nucleocapsid Proteins 0
envelope protein, SARS-CoV-2 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

20906

Subventions

Organisme : Hebei Medical University
ID : 2022007, USIP2021111
Organisme : Hebei Medical University
ID : 2022007, USIP2021111
Organisme : Hebei Medical University
ID : 2022007, USIP2021111
Organisme : Hebei Medical University
ID : 2022007, USIP2021111

Informations de copyright

© 2024. The Author(s).

Références

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Auteurs

Jiapei Hu (J)

Tangshan Clinical Medical College, Hebei Medical University, Tangshan, Hebei, China.
Graduate School, Hebei Medical University, Shijiazhuang, Hebei, China.

Jiayi Hu (J)

Tangshan Clinical Medical College, Hebei Medical University, Tangshan, Hebei, China.
Graduate School, Hebei Medical University, Shijiazhuang, Hebei, China.

Li Jin (L)

Tangshan Clinical Medical College, Hebei Medical University, Tangshan, Hebei, China.
Graduate School, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.

Dakang Hu (D)

Department of Laboratory Medicine, Taizhou Municipal Hospital, Taizhou, Zhejiang, China.

Philip K Nicholls (PK)

School of Medical, Molecular and Forensic Sciences, Murdoch University, 90 South Street, Murdoch, WA, 6149, Australia.

Tao Wang (T)

Telethon Kids Institute, Perth Children's Hospital, Nedlands, WA, Australia.
Medical School, University of Western Australia, Nedlands, WA, Australia.

Yonglin Ren (Y)

School of Agricultural Science, Murdoch University, Murdoch, WA, Australia.

Dailun Hu (D)

Department of Pathogenic Biology, Hebei Medical University, 361 Zhongshan East Road, Shijiazhuang, 050017, Hebei, China. 17300574@hebmu.edu.cn.

Bin Ma (B)

School of Medical, Molecular and Forensic Sciences, Murdoch University, 90 South Street, Murdoch, WA, 6149, Australia. B.Ma@murdoch.edu.au.
Centre for Healthy Aging, Health Futures Institute, Murdoch University, Murdoch, WA, Australia. B.Ma@murdoch.edu.au.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Virus › Virus à ARN › Virus à ARN à brin positif › Nidovirales › Coronaviridae › Coronavirus › Betacoronavirus › Virus du SRAS › SARS-CoV-2 › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Hybridation in situ › Hybridation fluorescente in situ › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN viral › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Maladies de l'appareil respiratoire › Infections de l'appareil respiratoire › Pneumopathie infectieuse › Pneumopathie virale › COVID-19 › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Cellules › Cellules épithéliales › Cellules HEK293 › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Phosphoprotéines › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines virales › Protéines virales structurales › Protéines de l'enveloppe virale › Protéines d'enveloppe des coronavirus › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN messager › Use of high-resolution fluorescence in situ...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines virales › Protéines virales structurales › Protéines nucléocapside › Protéines de la nucléocapside des coronavirus › Use of high-resolution fluorescence in situ...