Aceh Journal of Animal Science
Vol 6, No 3 (2021): November 2021

In silico SYBR® green-based qPCR primer design for virulent protein C gene of Edwardsiella tarda

Rian Ka Praja (Khon Kaen University)
Reny Rosalina (Unknown)



Article Info

Publish Date
10 Aug 2021

Abstract

Edwardsiella tarda infection in fish results in economic losses in aquaculture industries worldwide. Besides, this pathogen is also a zoonotic agent causing gastrointestinal disease in humans. Detection of virulence factors expressed by E. tarda is a key for molecular diagnosis. This study aimed to design SYBR®Green-based quantitative PCR (qPCR) primer for E. tarda virulent protein C (EvpC) gene. A sequence of EvpC with accession number AY424360.4:6500-6991 obtained from GenBank NCBI was selected as the basis for qPCR primer design. qPCR primers were designed using Primer3 online software. Further analysis related to the secondary structure of each primer was carried out using Beacon Designer Free Edition. Five pairs of qPCR primers were successfully designed with Primer3. Based on the results of Primer3 and Beacon Designer analysis, F primer pair 1 (5'-GAATCCACCGACGATAAGCACAAA-3') and R primer pair 1 (5'-GACACGCAGCACCGACATCA-3') were the most favorable primer set since this primer met the criteria in terms of length, melting temperature, GC content, and self complementarity. In addition, this primer set had the highest (closest to zero) -ΔG value of cross dimer, self dimer, and hairpin. It can be concluded that SYBR® Green-based qPCR primer set for EvpC has been successfully designed. However, laboratory optimization experiments are essential for this designed primer set.

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Journal Info

Abbrev

AJAS

Publisher

Subject

Description

Aceh Journal of Animal Science (Aceh J. Anim. Sci.) is an international peer-review journal published by Faculty of Marine and Fisheries, Syiah Kuala University in collaboration with Indonesian Society for Oceanology (ISOI) of Aceh Chapter and supported by Institute of Graduate Studies Syiah Kuala ...