Development of high-resolution melting (HRM) assay to differentiate the species of Shigella isolates from stool and food samples.


Journal

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

Informations de publication

Date de publication:
10 01 2022
Historique:
received: 07 09 2021
accepted: 23 12 2021
entrez: 11 1 2022
pubmed: 12 1 2022
medline: 3 3 2022
Statut: epublish

Résumé

Shigella species, a group of intracellular foodborne pathogens, are the main causes of bacillary dysentery and shigellosis in humans worldwide. It is essential to determine the species of Shigella in outbreaks and food safety surveillance systems. The available immunological and molecular methods for identifying Shigella species are relatively complicated, expensive and time-consuming. High resolution melting (HRM) assay is a rapid, cost-effective, and easy to perform PCR-based method that has recently been used for the differentiation of bacterial species. In this study, we designed and developed a PCR-HRM assay targeting rrsA gene to distinguish four species of 49 Shigella isolates from clinical and food samples and evaluated the sensitivity and specificity of the assay. The assay demonstrated a good analytical sensitivity with 0.01-0.1 ng of input DNA template and an analytical specificity of 100% to differentiate the Shigella species. The PCR-HRM assay also was able to identify the species of all 49 Shigella isolates from clinical and food samples correctly. Consequently, this rapid and user-friendly method demonstrated good sensitivity and specificity to differentiate species of the Shigella isolates from naturally contaminated samples and has the potential to be implemented in public health and food safety surveillance systems.

Identifiants

pubmed: 35013489
doi: 10.1038/s41598-021-04484-1
pii: 10.1038/s41598-021-04484-1
pmc: PMC8748861
doi:

Substances chimiques

DNA, Bacterial 0

Types de publication

Evaluation Study Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

473

Informations de copyright

© 2022. The Author(s).

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Auteurs

Babak Pakbin (B)

Institute for Life Technologies, University of Applied Sciences Western Switzerland Valais-Wallis, 1950 Sion 2, Sierre, Switzerland.
Department of Food Hygiene and Quality of Control, Faculty of Veterinary Medicine, University of Tehran, P.O. Box: 14155-6453, Tehran, Iran.

Afshin Akhondzadeh Basti (AA)

Department of Food Hygiene and Quality of Control, Faculty of Veterinary Medicine, University of Tehran, P.O. Box: 14155-6453, Tehran, Iran. aakhond@ut.ac.ir.

Ali Khanjari (A)

Department of Food Hygiene and Quality of Control, Faculty of Veterinary Medicine, University of Tehran, P.O. Box: 14155-6453, Tehran, Iran.

Wolfram Manuel Brück (WM)

Institute for Life Technologies, University of Applied Sciences Western Switzerland Valais-Wallis, 1950 Sion 2, Sierre, Switzerland.

Leila Azimi (L)

Pediatric Infections Research Center, Research Institute of Children's Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abdollah Karimi (A)

Pediatric Infections Research Center, Research Institute of Children's Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

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