The Potential of The ToxR Gene As A Detection Tool for Vibrio parahaemolyticus in Seafood
Muktiningsih Nurjayadi (a*), Ismaya Krisdawati (a), Gladys Indira Putri (a), Jefferson Lynford Declan (a), Dandy Akbar Juliansyah (a), Maharanianska Azzahra (a), Irvan Maulana (a), Atikah Nur Rahmawati (a), Irma Ratna Kartika (a), Fera Kurniadewi (a), Dalia Sukmawati (b), Sri Rahayu (b), Vira Saamia (c), Dwi Anna Oktaviani Saputro (c), I Made Wiranatha (c), H. E. Enshay (d)(e)(f)

a) Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj^ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia
*muktiningsih[at]unj.ac.id
b) Department of Biology, Faculty of Mathematics and Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj^ari, 9th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia
c) Center Forensic Laboratory of the Criminal Investigation, Police of the Republic of Indonesia, Ciapmbuan Babakan Madang, Bogor, 1681, Indonesia
d) Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
e) School of Chemical and Energy Engineering, Faculty of Engineering, Universiti Teknologi Malaysia (UTM), Skudai, Johor Bahru, Malaysia.
f) City of Scientific Research and Technology Applications, New Burg Al Arab, Alexandria, Egypt.

Abstract

Vibrio parahaemolyticus must be detected quickly and precisely. The purpose of this study is to use the Gradient Polymerase Chain Reaction (PCR) method to evaluate the primer that can produce an amplicon at the ideal temperature for the Vibrio parahaemolyticus toxR gene. The toxR primer pair could amplify Vibrio parahaemolyticus toxR fragments to form an amplicon with size 171 base pairs, according to the in silico analysis conducted for the primer design. Gradient PCR was used to assess the primer^s suitability for the design outcome and determine the ideal temperature to generate amplicons of the appropriate size with the temperature ranges from 53 to 62 degree Celsius. The purity (A260/280) of the Vibrio parahaemolyticus isolate used as a template is 1.92 and its concentration is 51 ng/ micro liter. The study^s findings kindly reveal that annealing temperatures of 60 degree Celsius produce the best outcomes for the toxR primer which results in the formation of a clear DNA band with a thickness band intensity and a size of 171 base pairs. Additionally, the primers and ideal temperatures discovered were utilized for the Real Time PCR stage in the creation of sensitive, targeted, and quicker tools for the detection of Vibrio parahaemolyticus in seafood.

Keywords: Foodborne Disease, Polymerase Chain Reaction, Seafood, toxR gene, Vibrio parahaemolyticus

Topic: Chemistry