Primers Design and PCR Optimization for Developing Salmonella Sp. Detection Method on Refillable Drinking Water Sample

DOI

10.2991/978-94-6463-166-1_62How to use a DOI?

Keywords

Refillable drinking water; Salmonella sp.; Primer Design; Gradient PCR

Abstract

One of the parameters of drinking water quality is not contaminated by coliform bacteria such as Salmonella sp. This bacteria can infect humans through oral routes causing infection of the gastrointestinal tract (Salmonellosis). Generally, analysis testing of coliform bacteria in water can be done using microbiological methods. However, microorganisms contained in refillable drinking water are usually in very small quantities and cause errors in sampling or in the calculation of the number of microorganisms. Testing with PCR technique can be used because it offers different advantages. PCR is specific, sensitive, fast and accurate to detect small amounts of target nucleic acids in drinking water samples. The study aims to obtain a Salmonella-specific PCR primer sequence for developing detection methods of refillable drinking water samples contaminated with Salmonella sp. And optimize the PCR condition using genomic DNA of Salmonella sp. as a template. Primer was designed using Geneious Prime and the specificity was checked using Primer BLAST on NCBI (National Center for Biotechnology Information). Meanwhile the PCR optimization was conducted using gradient PCR. The results of this study were Salmonella-specific PCR primer forward 5’-CCGTCTTATCTTGATTGAAGCCG-3’ and reverse 5’-CGTCATGATATTCCGCCCCA-3’ that amplified InvA gene of Salmonella sp. With amplicon size of 559 bp. The optimum PCR condition was the PCR program using annealing temperature at 59,5 ℃.

Copyright

© 2023 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Oswal Yuselman
AU  - Dwi Hilda Putri
AU  - Yuni Ahda
AU  - Afifatul Achyar
PY  - 2023
DA  - 2023/05/22
TI  - Primers Design and PCR Optimization for Developing Salmonella Sp. Detection Method on Refillable Drinking Water Sample
BT  - Proceedings of the 3rd International Conference on Biology, Science and Education (IcoBioSE 2021)
PB  - Atlantis Press
SP  - 521
EP  - 530
SN  - 2468-5747
UR  - https://doi.org/10.2991/978-94-6463-166-1_62
DO  - 10.2991/978-94-6463-166-1_62
ID  - Yuselman2023
ER  -