A Comparison Of Plating And Reverse Transcriptase Polymerase Chain Reaction Followed By Microchip Electrophoresis For The Inactivation Of Alicyclobacillus Acidoterrestris Using Saponin

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 μ-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. © 2014 Sociedade Brasileira de Química.2519197Eguchi, S.Y., Manfio, G.P., Pinhatti, M.E.M.C., Azuma, E., Variane, S.F., (1999) Acidothermophilic Sporeforming Bacteria (ATSB) in Orange Juices: Detection Methods, Ecology and Involvement in the Deterioration of Fruit Juices, 1. , 2nd ed.Fundação André Tosello & ABECitrus: CampinasEiroa, M., Junqueira, V., Schmidt, F., (1999) J. Food Protect., 62, p. 883Pettipher, G.L., Osmundson, M.E., Murphy, J.M., (1997) Lett. Appl. Microbiol., 24, p. 185Splittstoesser, D.F., Churey, J., Lee, C.Y., (1994) J. Food Protect., 57, p. 1080Yamazaki, K., Tekuda, H., Shinano, H., (1996) Biosci. Biotechnol. Biochem., 60, p. 543Orr, R.V., Shewefelt, R., Huang, C., Tefera, S., Beuchat, R.L., (2000) J. Food Protect., 63, p. 1517Knorr, D., (1995) New Methods of Food Preservation, 1. , 2nd ed.Blackie: LondonButz, P., (2002) Food Res. Internat., 35, p. 279Komitopoulou, E., Boziaris, I.S., Davies, E.A., Delves-Broughton, J., Radstrom, M.P., (1999) Int. J. Food Science Technol., 34, p. 81Yamazaki, K., Murakami, M., Kawai, Y., Inoue, N., Matsuda, T., (2000) Food Microbiology, 17, p. 315Grande, M.J., Lucas, R., Abriouel, H., Omar, N.B., Maqueda, M., Martínez-Bueno, M., Martínez-Cañamero, M., Gálvez, A., (2005) Int. J. Food Microbiology, 104, p. 289Minamikawa, M., Kawai, Y., Inoue, N., Yamazaki, K., (2005) Curr. Microbiology, 1, p. 22Falcone, P.M., Campaniello, D., Altieri, C., Sinigaglia, M., Corbo, M.R., Anese, M., Del Nobile, M.A., (2005) Ital. J. Food Sci., p. 142. , special issueBevilacqua, A., Corbo, M.R., Sinigaglia, M., (2008) Int. J. Food Sci. Technol., 43, p. 1271Hostettmann, K., Marston, A., (1995) Chemistry and Pharmacology of Natural Products, 1. , 3rd ed.Cambridge University Press: CambridgeFrancis, G., Kerem, Z., Makkar, H.P.S., Becker, K., (2002) Brit. J. Nutrit., 88, p. 587Sparg, S.G., Light, M.E., Van Staden, J., (2004) J. Ethnopharmacol., 94, p. 219Tsuzuki, J.K., Svidzinski, T.I.E., Shinobu, C.S., Silva, L.F.A., Rodrigues-Filho, E., Cortez, D.A.G., Ferreira, I.C.P., (2007) Anais Acad. Bras. Cienc., 79, p. 577Sheridan, G.E., Masters, C.I., Shallcross, J.A., MacKey, B.M., (1998) Appl. Environ. Microbiol., 64, p. 1313Williams, J.M., Trope, M., Caplan, D.J., Shugars, D.C., (2006) J. Endod., 32, p. 715McKillip, J.L., Jaykus, L.A., Drake, M., (1998) Appl. Environ. Microbiol., 12, p. 4264Nachamkin, I., Panaro, N., Huong Ung, M.L., Kricka, L., Wilding, P.J., (2001) J. Clin. Microbiol., 39, p. 754Gottwald, E., Müller, O., Polten, A., (2001) Electrophoresis, 22, p. 4016Jassona, V., Jacxsens, L., Luning, P., Rajkovic, A., Uyttendaele, M., (2010) Food Microbiol., 27, p. 710Yamazaki, K., Tekuda, H., Inoue, N., Shinano, H., (1996) Letters in Appl. Microbiology, 2, p. 350Alberice, J.V., Funes-Huacca, M.E., Guterres, S.B., Carrilho, E., (2012) Int. J. Food Microbioogy., 159, p. 130Panaro, N.J., Yuen, P.K., Sakazume, T., Fortina, P., Kricka, L.J., Wilding, P., (2000) Clin. Chem., 11, p. 1850Jabasini, M., Zhang, L., Dang, F., Xu, F., Almofli, M.R., Ewis, A.A., Lee, J., Baba, Y., (2002) Electrophoresis, 23, p. 1537Sodowich, I., Fadl, I., Burns, C., (2007) Electrophoresis, 28, p. 236