A comparison of plating and reverse transcriptase polymerase chain reaction followed by microchip electrophoresis for the inactivation of Alicyclobacillus acidoterrestris using saponin

Autor(es)

Juliana V. Alberice ^{[1, 2]} ; Maribel E. Funes-Huacca ^{[1, 2]} ; Emanuel Carrilho ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Chem Sao Carlos, BioMicS Lab, BR-13560970 Sao Carlos, SP - Brazil

^[2] Univ Campinas Unicamp, Natl Inst Sci & Technol Bioanalyt INCTBio, BR-13083970 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of the Brazilian Chemical Society; v. 25, n. 1, p. 91-97, 2014-01-00.

Resumo

It was used reverse-transcriptase polymerase chain reaction (RT-PCR) followed by capillary electrophoresis on a microchip to probe the viability of Alicyclobacillus acidoterrestris spores after inactivation with saponin and heat. The method was shown to be suitable for the purpose, and was faster and more sensitive than the traditional plating technique, the standard of the food industry. The limits of quantification and detection were 0.0107 and 0.0039 ng µL-1 of amplified DNA, respectively. The correlation coefficient (r) between traditional plating and our method was 0.9977, which indicates an excellent correspondence between them. Consequently, it was possible to assess, with confidence, the viability of bacteria using the RT-PCR reaction for detection. It was evaluated the potential of this molecular method over traditional microbiology in the inactivation of A. acidoterrestris by saponin as an effective agent, potentiated by heat. (AU)

• Processo FAPESP: 01/14391-4 - Identificação de Alicyclobacillus acidoterrestris em sucos de laranja pela reação em cadeia da polimerase por transcrição reversa (RT-PCR) e métodos eletroforéticos de análise
• Beneficiário: Emanuel Carrilho
• Modalidade de apoio: Auxílio à Pesquisa - Regular