Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology
Vol 15, No 2 (2020): August 2020

Detection of rtxA Gene as a Biomarker of Seafood-Borne Pathogen Vibrio cholerae using In Silico PCR Assay

Stalis Norma Ethica (Magister Program of Medical Laboratory Science, Universitas Muhammadiyah Semarang)
Nur Hidayati (Magister Program of Medical Laboratory Science, Universitas Muhammadiyah Semarang, Jl. Kedungmundu Raya No.18, Semarang, Indonesia 50273)
Hayatun Fuad (Magister Program of Medical Laboratory Science, Universitas Muhammadiyah Semarang, Jl. Kedungmundu Raya No.18, Semarang, Indonesia 50273)
Chaerul Arham (Politeknik Kesehatan Muhammadiyah Makassar, Jl. Ratulangi no. 101, Makassar, Indonesia, 90222)
Rivana Ariyadi (Sekolah Tinggi Ilmu Kesehatan Muhammadiyah Ciamis, Jl. K.H Ahmad Dahlan no. 20, Ciamis, Indonesia)
Ellyka Purwaningrum (Medical Faculty, Universitas Wahid Hasyim Semarang, Jl. Kolonel Warsito, Semarang, Indonesia,)
Kazi Mohammad Zillur Rahman (Centre for Marine Science and Innovation, University of New South Wales, Level 5 East, Biological Sciences North (D26))



Article Info

Publish Date
27 Aug 2020

Abstract

Seafood-borne outbreaks caused by Vibrio cholerae have led to the increased need for food safety risk assessment of marine products. An in silico investigation about the potential of virulence gene of V. cholerae, rtxA, as a DNA biomarker of the toxigenic bacterium has been carried out. The aim of this study was to use the bacterial DNA biomarker sequence as a tool to facilitate early rapid detection of cholera infection. Five specific pairs of primers were designed from the rtxA open reading frame DNA of V. cholerae O1 biovar El Tor str. N16961 genomic DNA using Primer3Plus. Next, in silico Polymerase Chain Reaction (PCR) assay was carried out using the newly designed primers and 25 genomic DNA of vibrio spp. retrieved from the in silico database. One of the five designed pairs of primers, RtxAOF-RtxAOR: ‘5-CGCAAAACAGTTTCAGCCGA-3’ and 5’-AGGTTGGTCTTTTGTGGCCA-3’, could result in single DNA amplicon sized 518 bp only from V. cholerae species. No amplicon bands were produced from 17 other vibrio genomes studied using similar RtxAF-RtxAR primers. A further check showed that the amplicon was indeed part of the rtxA gene of V. cholerae. Based on this in silico study, rtxA gene appeared to be a DNA biomarker of V. cholerae, which is potential to facilitate rapid diagnosis of the virulence bacterium using in silico PCR assay.

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Journal Info

Abbrev

squalen

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology

Description

Squalen publishes original and innovative research to provide readers with the latest research, knowledge, emerging technologies, postharvest, processing and preservation, food safety and environment, biotechnology and bio-discovery of marine and fisheries. The key focus of the research should be ...