Deteksi bakteri pembentuk amina biogenik pada ikan Scombridae secara multiplex PCR
Detection of Biogenic Amine Producing Bacteria in Scombridae Fish based on Multiplex PCR assay
Biogenic amines are nitrogenous base components formed by amino acid decarboxylation. Biogenic amines that are often detected in fishery products are histamine, tyramine and cadaverine. The amino acid decarboxylation coding genes are histidine decarboxylase (hdc), tyrosine decarboxylase (tdc) and lysine decarboxylase (ldc). The purpose of this study was to isolate DNA, select pre-enrichment media that can express biogenic amine forming genes using multiplex PCR , as well as identify biogenic amine forming bacteria. The specimens used were frozen tuna, little tuna, skipjack, and loin tuna. In addition, Indonesian traditional tuna sucha as pindang potong, pindang bumbu kuning were observed. Tuna stored at room temperature for 3 days was used as positive control. The study consisted of the pre-enrichment stage of bacteria on lactose broth and marine broth medias, DNA isolation (sample with and non pre-enrichment), purity test and isolate concentration of target gene of hdc, tdc, ldc using multiplex PCR. The result obtained were generally in a good quality of bacterial DNA isolates, with concentration ranged between 15.75-157.84 ng/μL. The multiplex PCR was successful in expressing hdc, tdc and ldc genes in scombridae fish on marine broth pre-enrichment media. The optimum condition for PCR amplification was annealing at 50°C for 90 seconds. Biogenic amine bacteria were identified, namely histamine forming bactria were Morganella morganii, Enterobacter aerogenes and Acinetobacter baumannii, cadaverin forming bacteria were Citrobacter sp., Hafnia paralvei, Obesumbacterium proteus Enterobacteer cloacae and Enterobacter hormachei, tyramine forming bacteria is Enterococcus faecalis.
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