Detecção molecular de Levilactobacillus brevis em cerveja por amplificação de DNA
DOI:
https://doi.org/10.18593/evid.37088Palavras-chave:
Cerveja, Diagnóstico, DNA, Indústria Cervejeira, Levilactobacillus brevisResumo
O setor cervejeiro brasileiro é um dos maiores empregadores do país e um pilar econômico significativo, responsável pela produção de bilhões de litros de cerveja anualmente. No entanto, enfrenta desafios relacionados à contaminação por BSMs (Beer Spoilage Microorganisms; ou microrganismos deteriorantes de cerveja), como Levilactobacillus brevis, que comprometem a qualidade do produto e geram prejuízos econômicos. Este trabalho teve como objetivo avaliar a eficiência de primers para a detecção específica de L. brevis a partir da aplicação das técnicas de Polimerase Chain Reaction (PCR) e Loop-Mediated Isothermal Amplification (LAMP). Cepas de Levilactobacillus brevis foram cultivadas em laboratório, além de outras bactérias ácido-lácticas dos gêneros Lentilactobacillus e Lactiplantibacillus, bem como bactérias heterólogas e fungos para os testes de especificidade. O DNA genômico de todos os microrganismos foi extraído e utilizado na composição de pools de DNA, utilizado nas análises de especificidade por PCR e LAMP. Para os testes de sensibilidade, o DNAg de L. brevis foi submetido a diluições seriadas e empregado nas técnicas de PCR, qPCR e LAMP. Os resultados da PCR demonstraram que os primers desenvolvidos apresentaram eficiência, porém razoável inespecificidade para Lentilactobacillus buchneri, outro contaminante de cerveja. O LAMP apresentou uma alternativa promissora à PCR, com alta especificidade e sensibilidade, amplificando até 100 fg/µL de DNAg. Este estudo reforça o potencial das metodologias moleculares, com destaque para o LAMP, e evidencia a necessidade de aperfeiçoamento contínuo dessas técnicas, a fim de assegurar soluções mais rápidas, precisas e eficazes para o controle de qualidade na indústria cervejeira.
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