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Journals / Gıda / 2021 / Cilt: 47 - Sayı: 1

RAPID DETECTION OF LISTERIA MONOCYTOGENES IN CHICKEN MEATS WITH COLORIMETRIC LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) METHOD

KOLORİMETRİK LOOP-MEDIATED İZOTERMAL AMPLİFİKASYON METODU İLE LISTERIA MONOCYTOGENES’İN TAVUK ETLERİNDE HIZLI TESPİTİ

Mehmet YÜKSEL
Selahattin SERT
Arzu KAVAZ YÜKSEL
Bülent ÇETİN
Mustafa GÜRSES
Journal
Gıda
Pages
121–135
Publication Date
2021-12-23
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

These results demonstrated the use of HNB-LAMP method for, as an alternative to PCR, sensitive and specific to L. monocytogenes under isothermal conditions, could use as a simple, rapid detection technique and has the potential for detection of L. monocytogenes from the chicken meats.

📌 3 Key Takeaways

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Objective & Hypothesis:The object of this study is to evaluate the performance of the colorimetric loop-mediated isothermal amplification (LAMP) in detecting Listeria monocytogenes from chicken meats (n: 50; whole wing, breast, drumstick; totally N: 150).
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Key Findings & Data:For this purpose, the chicken meats were artificially contaminated with 100-104 CFU/25 g (or sample) of L. monocytogenes and 5 others bacteria were used as competitive microbiota.
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Conclusion & Impact:These results demonstrated the use of HNB-LAMP method for, as an alternative to PCR, sensitive and specific to L. monocytogenes under isothermal conditions, could use as a simple, rapid detection technique and has the potential for detection of L. monocytogenes from the chicken meats.
🔬Design:Research Article
📍Scope:150 samples
⚙️Variables:Loop-mediated izotermal amplifikasyon (LAMP) · Gerçek zamanlı PZR · Listeria monocytogenes

Abstract

The object of this study is to evaluate the performance of the colorimetric loop-mediated isothermal amplification (LAMP) in detecting Listeria monocytogenes from chicken meats (n: 50; whole wing, breast, drumstick; totally N: 150). For this purpose, the chicken meats were artificially contaminated with 100-104 CFU/25 g (or sample) of L. monocytogenes and 5 others bacteria were used as competitive microbiota. After pre-enrichment, the samples were analyzed using conventional cultural, LAMP and real-time PCR methods. Primer sets of virulence gene hlyA were used for the L. monocytogenes-specific visualized LAMP with hydroxynaphthol blue (HNB) dye and amplified the target gene using specific primers at 65 °C for 45 min. As a result of analysis performed by three methods in the naturally contaminated samples, the presence of L. monocytogenes was detected in 9 of 150 samples (6%). The LAMP, qPCR and conventional method showed similar detection performances for L. monocytogenes in the naturally or artifically contaminated samples. These results demonstrated the use of HNB-LAMP method for, as an alternative to PCR, sensitive and specific to L. monocytogenes under isothermal conditions, could use as a simple, rapid detection technique and has the potential for detection of L. monocytogenes from the chicken meats.

Özet

Bu çalışmanın amacı tavuk etlerinden (n:50; bütün kanat, göğüs, baget; toplamda N: 150) Listeria monocytogenes’in tespitinde kolorimetrik loop-mediated izotermal amplifikasyon’un (LAMP) performansını değerlendirmektir. Bu amaçla, tavuk etleri 100-104 CFU/25 g (veya bütün tavuk eti) seviyede L. monocytogenes ve rekabetçi mikrobiyota olarak kullanılan 5 diğer bakteri (Salmonella Typhimurium, Campylobacter jejuni, Campylobacter coli, Staphylococcus aureus, Citrobacter freundii) ile inoküle edildi. Ön zenginleştirme sonrası örnekler geleneksel kültürel, gerçek zamanlı PZR ve LAMP kullanılarak analiz edildi. Virulans hlyA gen’in primer setleri (L. monocytogenes-özgü) hidroksinaftol mavisi (HNB) ile görselleştirilmiş LAMP için kullanıldı. Bu hedef gen 65°C 45 dakikada spesifik primerler kullanılarak çoğaltıldı. Üç metot ile gerçekleştirilen analizlerin sonucunda doğal olarak kontamine olmuş 150 örneğin 9’unda (%6) L. monocytogenes varlığı tespit edildi. LAMP, qPCR ve klasik metot doğal ve yapay olarak kontamine olmuş örneklerden L. monocytgones için aynı tespit performansını gösterdi. Bu sonuçlar HNB-LAMP yönteminin, PCR'a alternatif olarak, izotermal koşullar altında L. monocytogenes'e duyarlı, spesifik, basit, hızlı bir tespit tekniği olarak kullanılabileceğini ve L. monocytogenes'in tavuk etlerinden saptanma potansiyeline sahip olduğunu gösterdi.

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Cite This Article

YÜKSEL, M., SERT, S., YÜKSEL, A. K., ÇETİN, B., GÜRSES, M. (2021). RAPID DETECTION OF LISTERIA MONOCYTOGENES IN CHICKEN MEATS WITH COLORIMETRIC LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) METHOD. Gıda, Cilt: 47 - Sayı: 1, 121–135. https://ajindex.com/articles/394699

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References (6)

  1. Ayaz, N.D., Ayaz, Y., Kaplan, Y. Z., Dogru, A. K., & Aksoy, M. H. (2009). Rapid detection of Listeria monocytogenes, in chicken carcasses by IMS-PCR. Annals of Microbiology, 59(4), 741e744. Brescia, P. (2012). Micro-volume purity assessment of nucleic acids using A260/A280 ratio and spectral scanning. BioTek Instruments, Winooski, VT, Tech. Rep. AN060112–12, Rev, 06-04. da Costa, A., de Lira Nunes, M., Mendes-Marques, C., de Almeida, A. and Leal, N. (2014) Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Listeria monocytogenes and Major Pathogenic Serotypes. American Journal of Analytical Chemistry, 5, 1057-1064. doi: 10.4236/ajac.2014.516112. Feng, J., Dai, Z., Tian, X., & Jiang, X. (2018). Detection of Listeria monocytogenes based on combined aptamers magnetic capture and loop-mediated isothermal amplification. Food Control, 85, 443-452.
  2. Garrido-Maestu, A., Azinheiro, S., Carvalho, J., Abalde-Cela, S., Carbó-Argibay, E., Diéguez, L., et al. (2017). Combination of microfluidic loop-mediated isothermal amplification with gold nanoparticles for rapid detection of Salmonella spp. in food samples. Front. Microbiol. 8:2159. Goh, S. G., Kuan, C. H., Loo, Y. Y., Chang, W. S., Lye, Y. L., Soopna, P.,. & Son, R. (2012). Listeria monocytogenes in retailed raw chicken meat in Malaysia. Poultry science, 91(10), 2686-2690.
  3. Goto, M., Honda, E., Ogura, A., Nomoto, A., & Hanaki, K. I. (2009). Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques, 46(3), 167-172.
  4. Hitchins, A.D., Jinneman, K. and Chen, Y., 2017. BAM: Detection and enumeration of Listeria monocytogenes. Bacteriological Analytical Manual. Jamali, H., Paydar, M., Ismail, S., Looi, C. Y., Wong, W. F., Radmehr, B., et al. (2015). Prevalence, antimicrobial susceptibility and virulotyping of Listeria species and Listeria monocytogenes isolated from open-air fish markets. BMC Microbiology, 15(1), 144. Jin, Z., Ding, G., Li, G., Yang, G., Han, Y., Hao, N.,. & Li, W. (2020). Rapid detection of foodborne bacterial pathogens using visual high‐throughput microfluidic chip. Journal of Chemical Technology & Biotechnology, 95(5), 1460-1466.
  5. Kiddle, G., Hardinge, P., Buttigieg, N. et al. GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnol 12, 15 (2012). https://doi.org/10.1186/1472-6750-12-15
  6. Kreitlow, A., Becker, A., Ahmed, M., Kittl