Gıda Patojeni Salmonella enteritidis’in Hızlı Teşhisi için Aday Yeni Aptamer Çiftleri

Yıl 2024, Cilt: 40 Sayı: 1, 1 - 9, 30.04.2024

Dilek Çam Derin

Öz

Gıda patojenleri insan ve hayvan sağlığı için çok önemlidir. Enterik bir bakteri olan S. enteritidis en önemli patojenlerden biridir ve salmonellosise sebep olur. Kusma, ishal, ateş ve abdominal kramplar enfeksiyondan 12-72 saat sonra oluşan tipik semptomlardır ve ölüme sebep olabilir. ABD ve Avrupa’da gıda bazlı enfeksiyonlar öncelikle S. enteritidis’ten kaynaklanır. Bu nedenle bakterinin erken ve hızlı teşhisi her zaman oldukça önemlidir. Bu anlamda yanal akışlı testler (LFAs) hızlı sonuçlar alma ve pratik kullanım açısından oldukça avantajlıdır. LFA’ler yaygın olarak antikor ve aptamerler ile hazırlanır. Aptamerler tek zincirli DNA ve RNA molekülleridir ve hedef moleküle seçici şekilde bağlanır. Altın nanoparçacıklar (AuNPs) LFA’lerde en yaygın kullanılan işaretleyicilerdir. Bu çalışmada S. enteritidis’e özgün aptamer çiftleri kullanılarak aptamer çiftli strip test (APSA) platformu geliştirilmesi amaçlanmıştır. 1. aptamer çiftinin diğer çiftlerle karşılaştırıldığında S. enteritidis teşhisi için LFA geliştirilmesinde uygun olduğu gözlenmiştir ve M180 membran akış oranı ve pozitif sonuçlar açısından uygun membran olarak bulunmuştur.

Anahtar Kelimeler

S. enteritidis, Aptamer pairs, Lateral flow assay, Food pathogen

Proje Numarası

FF150219B05

New Aptamer Pairs as a Candidate for the Rapid Detection of Food Pathogen, Salmonella enteritidis

Öz

Food pathogens are crucial for human and animal health. S. enteritidis, enteric bacteria, is one of the most popular pathogen and causes salmonellosis. Vomitting, diarrhea, fever and abdominal cramps are typcial symptoms occuring at 12-72 hour after the infection and can cause to death. Food based infections in the USA and EU are primarily caused by S. enteritidis. Thus, early and rapid detection of bacteria is always highly significant. In this sense, lateral flow assays (LFAs) are notably advantageous with regards to the obtaining rapid results and practical use. LFAs are commonly prepared by antibodies and aptamers. Aptamers are single stranded DNA and RNA molecules and bind to the target molecules, selectively. Gold nanoparticles (AuNPs) are the most common labels used for LFAs. In this work, it is aimed to develop the aptamer paired strip assay (APSA) platform by using aptamer pairs specific to S. enteritidis. It was observed that 1st aptamer pairs are suitable for developing the LFAs for the recognition of S. enteritidis compared to the other pairs and M180 membrane was found as suitable membrane in terms of the flow rate and positive results.

Anahtar Kelimeler

S. enteritidis, Aptamer pairs, Lateral flow assay, Food pathogen

Proje Numarası

FF150219B05

Teşekkür

This work was supported by the Scientific Research Project at Çankırı Karatekin University (BAP- FF150219B05). The author acknowledges to Nanobiz NanoBiotechnological Systems R&D Limited, Dr. Ceren Bayrac (Karamanoğlu Mehmet Bey University, Department of Biomedical Engineering) and Dr. Mine TÜRKTAŞ ERKEN (Gazi University, Department of Biology) for their kindly help.

Kaynakça

• Levy, B. S., McIntire, W. 1974. The economic impact of a food-borne salmonellosis outbreak. JAMA, 230:1281-1282.
• EFSA and ECDC. 2018. The European Union One Health 2018 Zoonoses Report. EFSA Journal 2019;17(12):5926.
• ECDC. 2016. Salmonellosis- Annual Epidemiological Report 2016 (2014 data). Annual Epidemiological Report on Communicable Diseases in Europe. https://www.ecdc.europa.eu/en/publications-data/salmonellosis-annual-epidemiological-report-2016-2014-data (Date accessed: 31.01.2024).
• ECDC-EFSA. 2020. RAPID OUTBREAK ASSESSMENT, Multi-country outbreak of Salmonella Enteritidis infections linked to eggs, Third update. https://www.ecdc.europa.eu/en/publications-data/multi-country-outbreak-salmonella-enteritidis-infections-linked-eggs (Date accessed: 31.01.2024).
• MinLee, K., Runyon, M., Herrman, T. J., Phillips, R., Hsieh, J. 2015. Review of Salmonella detection and identification methods: Aspects of rapid emergency response and food safety. Food Control, 47: 264-276.
• Pathmanathan, S. G., Cardona-Castro, N., Sánchez-Jiménez, M. M., Correa-Ochoa, M. M., Puthucheary, S.D., Thong, K. L. 2003. Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene. Journal of Medical Microbiology, 52 (9): 773-776.
• Wen, C. Y., Hu, J., Zhang, Z. L., Tiani, Z. Q., Ou, G.P., Liao, Y. L, & Pang, D. W. 2013. One-step sensitive detection of Salmonella typhimurium by coupling magnetic capture and fluorescence identification with functional nanospheres. Analytical chemistry, 85(2): 1223-1230.
• Zhang, D., and Alocilja, E. C. 2008. Characterization of nanoporous silicon-based DNA biosensor for the detection of Salmonella Enteritidis. Sensors Journal, IEEE, 8(6): 775-780.
• Laderman, E. I., Whitworth, E., Dumaual, E., Jones, M., Hudak, A., Hogrefe, W., & Groen, J. 2008. Rapid, sensitive, and specific lateral-flow immunochromatographic point-of-care device for detection of herpes simplex virus type 2-specific immunoglobulin G antibodies in serum and whole blood. Clinical and Vaccine Immunology, 15(1): 159-163.
• He, Y., Zhang, S., Zhang, X., Baloda, M., Gurung, A. S., Xu, H., & Liu, G. 2011. Ultrasensitive nucleic acid biosensor based on enzyme–gold nanoparticle dual label and lateral flow strip biosensor. Biosensors and Bioelectronics, 26(5): 2018-2024.
• Deng, Y. P., Zhao, H. Q., Jiang, L. 2000. Applications of nanogold particles in biomimetic engineering. China Basic Science, 9: 11–17.
• Siciliano, G., Alsadig, A., Chiriacò, M. S., Turco, A., Foscarini, A., Ferrara, F., Gigli, G., Primiceri, E. 2024. Beyond traditional biosensors: Recent advances in gold nanoparticles modified electrodes for biosensing applications. Talanta, 268: 125280.
• Bayraç, C., Eyidoğan, F., Öktem, H. A. 2017. DNA aptamer-based colorimetric detection platform for Salmonella Enteritidis. Biosensors and Bioelectronics, 98; 22–28.
• Ellington, A. D., Szostak, J. W. 1990. In vitro selection of RNA molecules that bind specific ligands. Nature, 346(6287):818-22.
• Çam Derin, D., Gültekin, E., Gündüz, Elif., Otlu, B. 2024. Comparison of 6 aptamer-aptamer pairs on rapid detection of SARS-CoV-2 by lateral flow assay. Journal of AOAC International, DOI: 10.1093/jaoacint/qsae004 (in press).
• Zeng, H., Guo, W., Liang, B., Li, J., Zhai, X., Song, C., Zhao, W., Fan, E., Liu, Q. 2016. Self-paired monoclonal antibody lateral flow immunoassay strip for rapid detection of Acidovorax avenae subsp citrull., Analytical Bioanalytical Chemistry, 408(22): 6071–6078.
• Chen, A., Yang, S. 2015. Replacing antibodies with aptamers in lateral flow immunoassay, Biosensors and Bioelectronics, 71: 230–242.
• Fang, Z., Wu, W., Lu, X., Zeng, L. 2014. Lateral flow biosensor for DNA extraction-free detection of salmonella based on aptamer mediated strand displacement amplification Biosensors and Bioelectronics, 56, 192–197.
• Park, H. C., Baig, I.A., Lee, S. C., Moon, J. Y., Yoon, M. Y. 2014. Development of ssDNA Aptamers for the Sensitive Detection of Salmonella typhimurium and Salmonella enteritidis. Applied Biochemistry and Biotechnology, 174(2):793-802.
• Jin, L., Wang, S., Shao, Q., Cheng, Y. 2022. A rapid and facile analytical approach to detecting Salmonella Enteritidis with aptamer-based surface-enhanced Raman spectroscopy. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 267,120625.
• Huo, T. M., Peng, C. F., Xu, C. L., Liu, L. Q. 2006. Development of colloidalgold-based immunochromatographic assay for the rapid detection of medroxyprogesterone acetate residues. Food and Agricultural Immunology, 17:183-190.
• Kaiser, L., Weisser, J., Kohl, M., & Deigner, H. P. 2018. Small molecule detection with aptamer based lateral flow assays: Applying aptamer-C-reactive protein cross-recognition for ampicillin detection. Scientific Reports, 8: 5628.
• Zhang, G., Zhu, C., Huang, Y., Yan, J., Chen, A. 2018. A Lateral Flow Strip Based Aptasensor for Detection of Ochratoxin A in Corn Samples. Molecules, 23: 291.
• Cam, D., Oktem, H. A. 2019. Development of rapid dipstick assay for food pathogens, Salmonella, by optimized parameters. Journal of Food Science and Technology, 56(1):140–148.
• Shen, G., Zhang. S., Hu. X. 2013. Signal enhancement in a lateral flow immunoassay based on dual gold nanoparticle conjugates. Clinical Biochemistry, 46: 1734–1738.
• Li, J., Jing, L., Song, Y., Zhang, J., Chen, Q., Wang, B., Xia, X., Han, Q. 2018. Rapid Detection of Rongalite via a Sandwich Lateral Flow Strip Assay Using a Pair of Aptamers. Nanoscale Research Letters, 13: 296.
• Bua, T., Yaoa, X., Huangb, L., Doua, L., Zhaoa, B., Yanga, B., Lic, T., Wang J., Zhanga, D. .2020. Dual recognition strategy and magnetic enrichment based lateral flow assay toward Salmonella enteritidis detection. Talanta, 206: 120204.
• Kim, E., Yang, S. M., Kim, H. Y. 2023. Development of rapid on-site detection of Salmonella Enteritidis, S. Typhimurium, and S. Thompson in food samples using an ultrafast PCR system. Food Bioscience, 56, 103242.
• Zambry, N. S., Najib, M. A., Awang, M. S., Selvam, K., Khalid, M. F., Bustami, Y., Hamzah, H. H., Ozsoz, M., Manaf, A. A., Aziah, I. 2022. Aptamer-Based Electrochemical Biosensors for the Detection of Salmonella: A Scoping Review. Diagnostics (Basel), 12(12): 3186.