Articles Vol. 63 No. Số tiếng Anh (2022) 08/12/2022

ESTABLISHMENT OF MULTIPLEX PCR ASSAY FOR SIMULTANEOUS DETECTION OF SALMONELLA SPP., SHIGELLA SPP. AND ENTAMOEBA HISTOLYTICA

Le Thi Thuy1, Chu Van Men1, Nguyen Thi Ngoc Anh1, Tran Thi Loan1, Luu Thuy Anh1, Dao Thuy Trang1, Hoang Thu Trang1, Ho Huu Tho1, Ninh Thi Kim Oanh2
1 Military Medical University
2 Department of Foreign Languages Teaching, Hanoi Medical University
Corresponding author: hohuutho@vmmu.edu.vn
DOI: 10.52163/yhc.v63i9.554
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Keywords
Multiplex PCR, Semi-Nested PCR, Salmonella spp., Shigella spp., Entamoeba histolytica.
Abstract

Objective: To establish a novel multiplex PCR assay for simultaneous detection
of foodborne pathogens, including Salmonella spp., Shigella spp., and Entamoeba
histolytica, with high sensitivity, specificity, cost-effectiveness and time-saving.
Method: Multiplex PCR, Semi-Nested PCR and melting analysis for specifically
amplifying and distinguishing three highly conserved target gene regions (invA,
ipaH, 18S rRNA), in order to simultaneously detect Salmonella spp., Shigella spp.,
Entamoeba histolytica in a single reaction tube. Specific pathogens are characterized
based on the melting profiles of the PCR products collected on a realtime PCR
instrument following the second round of amplification.
Results: The established multiplex PCR assay is capable of simultaneously detecting
Salmonella spp., Shigella spp., and Entamoeba histolytica at a limit of detection of
5.86 copies/µl, 24.4 copies/µl và 53.6 copies/µl, respectively (confidence interval of
95%), the specificity reached 100% when evaluated with multiple microbial species
provided by the Department of Microbiology, 103 Military Hospital.
Conclusion: The novel multiplex PCR assay for simultaneous detection of Salmonella
spp., Shigella spp., Entamoeba histolytica has been successfully established.

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