Application of end-point PCR technique to detect bacteria encoding tyrosine decarboxylase (TDC) gene in scombridae fish

摘要

Tuna, little tuna, and skipjack can form tyramine from decarboxylation of tyrosine by tyrosine decarboxylase (TDC) enzyme. High tyramine level in fish could cause toxicological effects. Early detection of the tyramine-forming bacteria by using DNA-based methods is needed for seafood safety assurance because it is more sensitive, specific, and faster. This study was aimed to obtain bacterial isolates, detection of TDC and 16S rRNA genes and identifying species of bacteria that encode TDC and 16S rRNA genes. The methods of this study included several steps including bacterial cultivation, DNA isolation and amplification of TDC and 16S rRNA gene markers as well as sequencing of amplicons. Bacterial DNA isolates were successfully obtained from samples by cultivation method. The TDC gene was successfully amplified and identified as belong to Carnobacterium genus, whereas the 16S rRNA gene belong to Enterobacter tabaci, E. hormaechei, Escherichia marmotae, and Peptoniphilus genus. The DNA-based method targeted tyrosine decarboxylase gene can be applied for early detection of biogenic amine accumulation in fishery products.