Listeria monocytogenes Specific Detection Employing Novel Primers with In Silico Study to Detect Listeriolysin O Protein-Coding Gene hly

DOI

10.2991/978-94-6463-457-0_6How to use a DOI?

Keywords

Systemic Lupus Erythematosus (SLE); Hemoglobin Level; Leukocyte Count; ESR

Abstract

Listeria monocytogenes is a pathogenic bacterium, which prevalence in the aquatic ecosystem and seafood products become a major threat to public health. Several virulence genes had been targeted for the PCR (polymerase chain reaction) detection of L.monocytogenes including hly coding for listeriolysin O precursor, which resulted in many reported primers. This paper presents newly paired primers for selective PCR detection of L.monocytogenes strains based on hly sequence by in silico approach using Primer3Plus and In Silico Programs. The quality of the newly paired primers was compared to the previously reported ones from various studies using Autodimer program. As result, two paired primers HlyOF: 5'-GACAAATGTGCCGCCAAGAA-3' and ELMHLYR: 5'-GCGCTTGCAACTGCTCTTTA-3', as well as lmo0202_q_F: 5'-CTCCGCCTGCAAGTCCTAAG-3' and HlyR: 5'-CTTCACTGATTGCGCCGAAG-3' were found to be the best in performance in virtual PCR compared to other previously reported pairs of primers. Using these primers, positive 596-bp and 594-bp PCR products, respectively, could be amplified from each of all genome sequences of L.monocytogenes strains (41 strains) available in the database of In Silico Program, but not from those of other Listeria species. Compared to previously reported primers, the newly paired primers have better theoretical quality based on Tm difference. The F: 5'-GACAAATGTGCCGCCAAGAA-3' primer is a novelty as it has not been reported anywhere and named as HlyO. As a conclusion, this in silico study confirmed that hly gene is an apparent marker gene, which only present in L.monocytogenes, but absent in other Listeria species. Hence, the newly paired primers are the potential to be used as a selective PCR detection component of L.monocytogenes of various strains.

Copyright

© 2024 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  - Ragil Saptaningtyas
AU  - Stalis Norma Ethica
AU  - Chaerul Arham
AU  - Wikanastri Hersoelistyorini
AU  - Rivana Ariadi
AU  - Ayu Rahmawati Sulistyaningtyas
AU  - M. Ardi Afriansyah
AU  - Aditya Rahman Ernanto
PY  - 2024
DA  - 2024/07/22
TI  - Listeria monocytogenes Specific Detection Employing Novel Primers with In Silico Study to Detect Listeriolysin O Protein-Coding Gene hly
BT  - Proceedings of the 2nd Lawang Sewu International Symposium on Health Sciences: Medical Laboratory Technology (LSISHS-MLT 2023)
PB  - Atlantis Press
SP  - 45
EP  - 53
SN  - 2468-5747
UR  - https://doi.org/10.2991/978-94-6463-457-0_6
DO  - 10.2991/978-94-6463-457-0_6
ID  - Saptaningtyas2024
ER  -