RAPID DETECTION OF INVA GENES IN SALMONELLA ATCC BY LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP)
AbstractSalmonella spp. is a bacterium that causes typhoid salmonellosis (typhoid fever). These bacteria enter through the oral route, usually by contaminating food and drinks. Salmonella spp. has an invA gene that causes pathogenicity in humans. The culture method is used as a gold standard to detect Salmonella spp. However, this method requires infrastructure and is laborious. The purpose of this study is to develop detection of Salmonella spp. using the Loop-Mediated Isothermal Amplification (LAMP) method. The stages of the study began with the primary design of LAMP, Salmonella ATCC culture, DNA Isolation of Salmonella ATCC culture, and Salmonella ATCC LAMP Optimization. Primary LAMP that has been designed consists of three pairs. Salmonella ATCC DNA that has been isolated has a concentration of 4,5 ng/ul. The LAMP method shows positive results in detecting the InvA gene in Salmonella ATCC. Positive results are indicated by fluorescence in Salmonella ATCC DNA samples, whereas the negative results are shown in the absence of fluorescence in nuclease free water. The conclusion from the research that has been done, LAMP method can detect InvA gene in Salmonella ATCC.
Bugarel, M., Tudor, A., Loneragan, G. H., & Nightingale, K. K. (2017). Molecular detection assay of five Salmonella serotypes of public interest: Typhimurium, Enteritidis, Newport, Heidelberg, and Hadar. Journal of microbiological methods, 134, 14-20.doi:10.1016/j.mimet.2016.12.011
De Jong, Hanna K., Chris M. Parry, Tom van der Poll, and W. Joost Wiersinga. 2012. Host–Pathogen Interaction in Invasive Salmonellosis. PLoS Pathogens 8(10)
Fan, F., Du, P., Kan, B., Yan, M., 2015. The Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Salmonella enterica serovar Typhi. PLOS ONE 10, e0124507. doi:10.1371/journal.pone.0124507
Hara-Kudo, Y., Yoshino, M., Kojima, T., & Ikedo, M. (2005). Loop-mediated isothermal amplification for the rapid detection of Salmonella. FEMS microbiology letters, 253(1), 155-161.
Kokkinos, P. A., Ziros, P. G., Bellou, M., & Vantarakis, A. (2014). Loop-mediated isothermal amplification (LAMP) for the detection of Salmonella in food. Food Analytical Methods, 7(2), 512-526.
Li, Y., Fan, P., Zhou, S., & Zhang, L. (2017). Loop-mediated isothermal amplification (LAMP): a novel rapid detection platform for pathogens. Microbial pathogenesis, 107, 54-61.
Pham, O.H., McSorley, S.J., 2015. Protective host immune responses to Salmonella infection. Future Microbiol. 10, 101–110. doi:10.2217/fmb.14.98
Sambrook, J., Russell, D. W., Janssen, K., & Argentine, J. (2001). Molecular cloning: a laboratory manual on the web. Cold Spring Harbor Laboratory.
Sun, Y., Quyen, T. L., Hung, T. Q., Chin, W. H., Wolff, A., & Bang, D. D. (2015). A lab-on-a-chip system with integrated sample preparation and loop-mediated isothermal amplification for rapid and quantitative detection of Salmonella spp. in food samples. Lab on a Chip, 15(8), 1898-1904.
Wilson, K. (2001). Preparation of genomic DNA from bacteria. Current protocols in molecular biology, 56(1), 2-4.
Wu, G.P., Chen, S.H., Levin, R.E., 2015. Application of ethidium bromide monoazide for quantification of viable and dead cells of Salmonella enterica by real-time loop-mediated isothermal amplification. J. Microbiol. Methods 117, 41–48.doi:10.1016/j.mimet.2015.07.012
Zhao, X., Lin, C. W., Wang, J., & Oh, D. H. (2014). Advances in rapid detection methods for foodborne pathogens. J. Microbiol. Biotechnol, 24(3), 297-312.
Zhuang, L., Gong, J., Li, Q., Zhu, C., Yu, Y., Dou, X., Liu, X., Xu, B., Wang, C., 2014. Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene. Lett. Appl. Microbiol. 59, 658–664. doi:10.1111/lam.12328
Copyright (c) 2019 Jurnal Ilmiah Ibnu Sina (JIIS): Ilmu Farmasi dan Kesehatan
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.