Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Rapid and Sensitive Detection of Salmonella in Chickens Using Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Dipstick

  • Liu, Zhi-Ke (College of Animal Science and Technology, Shihezi University) ;
  • Zhang, Qiu-Yu (College of Animal Science and Technology, Henan Institute of Science and Technology) ;
  • Yang, Ning-Ning (College of Animal Science and Technology, Shihezi University) ;
  • Xu, Ming-Guo (College of Animal Science and Technology, Shihezi University) ;
  • Xu, Jin-Feng (College of Life Sciences, Shihezi University) ;
  • Jing, Ming-Long (College of Animal Science and Technology, Shihezi University) ;
  • Wu, Wen-Xing (College of Animal Science and Technology, Shihezi University) ;
  • Lu, Ya-Dong (College of Animal Science and Technology, Shihezi University) ;
  • Shi, Feng (College of Life Sciences, Shihezi University) ;
  • Chen, Chuang-Fu (College of Animal Science and Technology, Shihezi University)
  • Received : 2017.12.06
  • Accepted : 2018.03.21
  • Published : 2019.03.28
Download PDF
⟨ Previous Next ⟩

Abstract

Salmonellosis is a highly contagious bacterial disease that threatens both human and poultry health. Tests that can detect Salmonella in the field are urgently required to facilitate disease control and for epidemiological investigations. Here, we combined loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) to rapidly and accurately detect Salmonella. LAMP primers were designed to target the Salmonella invA gene. LAMP conditions were optimized by adjusting the ratio of inner to outer primers, $MgSO_4$ concentration, dNTP mix concentration, amplification temperature, and amplification time. We evaluated the specificity of our novel LAMP-LFD method using six Salmonella species and six related non-Salmonella strains. All six of the Salmonella strains, but none of the non-Salmonella strains, were amplified. LAMP-LFD was sensitive enough to detect concentrations of Salmonella enterica subsp. enterica serovar Pullorum genomic DNA as low as $89fg/{\mu}l$, which is 1,000 times more sensitive than conventional PCR. When artificially contaminated feed samples were analyzed, LAMP-LFD was also more sensitive than PCR. Finally, LAMP-LFD gave no false positives across 350 chicken anal swabs. Therefore, our novel LAMP-LFD assay was highly sensitive, specific, convenient, and fast, making it a valuable tool for the early diagnosis and monitoring of Salmonella infection in chickens.

Keywords

References

  1. Tang F, Pang DW, Chen Z, Shao JB, Xiong LH, Xiang YP, et al. 2016. Visual and efficient immunosensor technique for advancing biomedical applications of quantum dots on Salmonella detection and isolation. Nanoscale 8: 4688-4698. https://doi.org/10.1039/C5NR07424J
  2. Lahiri C, Pawar S, Sabarinathan R, Ashraf MI, Chand Y, Chakravortty D. 2014. Interactome analyses of Salmonella pathogenicity islands reveal SicA indispensable for virulence. J. Theor. Biol. 363: 188-197. https://doi.org/10.1016/j.jtbi.2014.08.013
  3. Vohra P, Bugarel M, Turner F, Loneragan GH, Hope JC, Hopkins J, et al. 2017. Quantifying the survival of multiple Salmonella enterica serovars in vivo using massively-parallel whole genome sequencing to predict zoonotic risk. Appl. Environ. Microbiol. 84: e02262-17.
  4. Ji Y, Li W, Zhang Y, Chen L, Zhang Y, Zheng X, et al. 2017. QseB mediates biofilm formation and invasion in Salmonella enterica serovar Typhi. Microb. Pathog. 104: 6-11. https://doi.org/10.1016/j.micpath.2017.01.010
  5. Patterson PH, Venkitanarayanan K, Kariyawasam S. 2014. Introduction: reducing Salmonella Enteritidis contamination of shell eggs. J. Appl. Poultry Res. 23: 323-329. https://doi.org/10.3382/japr.2014-00940
  6. Fatica MK, Schneider KR. 2011. Salmonella and produce: survival in the plant environment and implications in food safety. Virulence 2: 573-579. https://doi.org/10.4161/viru.2.6.17880
  7. Rehkopf AC, Byrd JA, Coufal CD, Duong T. 2017. Advanced oxidation process sanitization of hatching eggs reduces Salmonella in broiler chicks. Poult. Sci. 96: 3709-3716. https://doi.org/10.3382/ps/pex166
  8. Braden CR. 2006. Salmonella enterica serotype Enteritidis and eggs: a national epidemic in the United States. Clin. Infect. Dis. 43: 512-517. https://doi.org/10.1086/505973
  9. Kim SY, Lee SK, Park MS, Na HT. 2016. Analysis of the fluoroquinolone antibiotic resistance mechanism of Salmonella enterica isolates. J. Microbiol Biotechnol. 26: 1605-1612. https://doi.org/10.4014/jmb.1602.02063
  10. Aribam SD, Ogawa Y, Matsui H, Hirota J, Okamura M, Akiba M, et al. 2015. Monoclonal antibody-based competitive enzyme-linked immunosorbent assay to detect antibodies to O:4 Salmonella in the sera of livestock and poultry. J. Microbiol. Methods 108: 1-3. https://doi.org/10.1016/j.mimet.2014.11.002
  11. Nde CW, Fakhr MK, Doetkott C, Logue CM. 2008. An evaluation of conventional culture, invA PCR, and the realtime PCR iQ-Check kit as detection tools for Salmonella in naturally contaminated premarket and retail turkey. J. Food Prot. 71: 386-391. https://doi.org/10.4315/0362-028X-71.2.386
  12. Dong J, Zhao H, Xu M, Ma Q, Ai S. 2013. A label-free electrochemical impedance immunosensor based on AuNPs/PAMAM-MWCNT-Chi nanocomposite modified glassy carbon electrode for detection of Salmonella Typhimurium in milk. Food Chem. 141: 1980-1986. https://doi.org/10.1016/j.foodchem.2013.04.098
  13. Bell RL, Jarvis KG, Ottesen AR, McFarland MA, Brown EW. 2016. Recent and emerging innovations in Salmonella detection: a food and environmental perspective. Microb. Biotechnol. 9: 279-292. https://doi.org/10.1111/1751-7915.12359
  14. Ohud MAH, Hayam SA, And EMH. 2015. Molecular typing of Salmonella enterica serovars Typhimurium and Enteritidis isolated from Taif area of Saudi Arabia by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Afr. J. Microbiol. Res. 9: 651-661. https://doi.org/10.5897/AJMR2014.7292
  15. Oh SY, Heo NS, Shukla S, Cho HJ, Vilian ATE, Kim J, et al. 2017. Development of gold nanoparticle-aptamer-based LSPR sensing chips for the rapid detection of Salmonella typhimurium in pork meat. Sci. Rep. 7: 10130. https://doi.org/10.1038/s41598-017-10188-2
  16. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28: E63. https://doi.org/10.1093/nar/28.12.e63
  17. Haridas DV, Pillai D, Manojkumar B, Nair CM, Sherief PM. 2010. Optimisation of reverse transcriptase loop-mediated isothermal amplification assay for rapid detection of Macrobrachium rosenbergii noda virus and extra small virus in Macrobrachium rosenbergii. J. Virol. Methods 167: 61-67. https://doi.org/10.1016/j.jviromet.2010.03.011
  18. Techathuvanan C, Draughon FA, D'Souza DH. 2010. Loopmediated isothermal amplification (LAMP) for the rapid and sensitive detection of Salmonella Typhimurium from pork. J. Food Sci. 75: M165-172. https://doi.org/10.1111/j.1750-3841.2010.01554.x
  19. Lalle M, Possenti A, Dubey JP, Pozio E. 2018. Loopmediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect toxoplasma gondii oocyst in readyto-eat salad. Food Microbiol. 70: 137-142. https://doi.org/10.1016/j.fm.2017.10.001
  20. Tsai SM, Liu HJ, Shien JH, Lee LH, Chang PC, Wang CY. 2012. Rapid and sensitive detection of infectious bursal disease virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick. J. Virol. Methods 181: 117-124. https://doi.org/10.1016/j.jviromet.2011.09.002
  21. Makaraviciute A, Jackson CD, Millner PA, Ramanaviciene A. 2016. Considerations in producing preferentially reduced half-antibody fragments. J. Immunol. Methods 429: 50-56. https://doi.org/10.1016/j.jim.2016.01.001
  22. Ge Y, Wu B, Qi X, Zhao K, Guo X, Zhu Y, et al. 2013. Rapid and sensitive detection of novel avian-origin influenza A (H7N9) virus by reverse transcription loop-mediated isothermal amplification combined with a lateral-flow device. PLoS One 8: e69941. https://doi.org/10.1371/journal.pone.0069941
  23. Huang HL, Zhu P, Zhou CX, He S, Yan XJ. 2017. The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Karlodinium veneficum. Harmful Algae 62: 20-29. https://doi.org/10.1016/j.hal.2016.11.022
  24. Puthawibool T, Senapin S, Flegel TW, Kiatpathomchai W. 2010. Rapid and sensitive detection of Macrobrachium rosenbergii nodavirus in giant freshwater prawns by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick. Mol. Cell. Probes 24: 244-249. https://doi.org/10.1016/j.mcp.2010.07.003
  25. Kiatpathomchai W, Jaroenram W, Arunrut N, Jitrapakdee S, Flegel TW. 2008. Shrimp Taura syndrome virus detection by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick. J. Virol. Methods 153: 214-217. https://doi.org/10.1016/j.jviromet.2008.06.025
  26. Kaewphinit T, Arunrut N, Kiatpathomchai W, Santiwatanakul S, Jaratsing P, Chansiri K. 2013. Detection of Mycobacterium tuberculosis by using loop-mediated isothermal amplification combined with a lateral flow dipstick in clinical samples. Biomed. Res. Int. 2013: 926230.
  27. Mahittikorn A, Thammasonthijarern N, Roobthaisong A, Udonsom R, Popruk S, Siri S, et al. 2017. Development of a loop-mediated isothermal amplification technique and comparison with quantitative real-time PCR for the rapid visual detection of canine neosporosis. Parasit Vectors 10: 394. https://doi.org/10.1186/s13071-017-2330-2
  28. Lee N, Kwon KY, Oh SK, Chang HJ, Chun HS, Choi SW. 2014. A multiplex PCR assay for simultaneous detection of Escherichia coli O157:H7, Bacillus cereus, Vibrio parahaemolyticus, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus in Korean ready-to-eat food. Foodborne Pathog. Dis. 11: 574-580. https://doi.org/10.1089/fpd.2013.1638
  29. Wang F, Jiang L, Ge B. 2012. Loop-mediated isothermal amplification assays for detecting shiga toxin-producing Escherichia coli in ground beef and human stools. J. Clin. Microbiol. 50: 91-97. https://doi.org/10.1128/JCM.05612-11
  30. Wang X, Ying S, Wei X, Yuan J. 2017. Development of a gold nanoparticle-based universal oligonucleotide microarray for multiplex and low-cost detection of foodborne pathogens. Int. J. Food Microbiol. 253: 66-74. https://doi.org/10.1016/j.ijfoodmicro.2017.05.005
  31. Wu GP, Chen SH, Levin RE. 2015. Rapid real-time loopmediated isothermal amplification combined with coated activated carbon for detection of low numbers of Salmonella enterica from lettuce without enrichment. Food Control. 56: 47-52. https://doi.org/10.1016/j.foodcont.2015.03.008
  32. Yamazaki W, Taguchi M, Kawai T, Kawatsu K, Sakata J, Inoue K, et al. 2009. Comparison of loop-mediated isothermal amplification assay and conventional culture methods for detection of Campylobacter jejuni and Campylobacter coli in naturally contaminated chicken meat samples. Appl. Environ. Microbiol. 75: 1597-1603. https://doi.org/10.1128/AEM.02004-08
  33. Yang Q, Domesle KJ, Wang F, Ge B. 2016. Rapid detection of Salmonella in food and feed by coupling loop-mediated isothermal amplification with bioluminescent assay in realtime. BMC Microbiol. 16: 112. https://doi.org/10.1186/s12866-016-0730-7
  34. Higuchi R, Fockler C, Dollinger G, Watson R. 1993. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology (N Y) 11: 1026-1030.
  35. Ahern H. Convenient, performance-boosting products put PCR on the fast track-the scientist-magazine of the life sciences. Scientist 10.
  36. Xu G, Hu L, Zhong H, Wang H, Yusa S, Weiss TC, et al. 2012. Cross priming amplification: mechanism and optimization for isothermal DNA amplification. Sci. Rep. 2: 246. https://doi.org/10.1038/srep00246
  37. Barbour EK, Ayyash DB, Alturkistni W, Alyahiby A, Yaghmoor S, Iyer A, et al. 2015. Impact of sporadic reporting of poultry Salmonella serovars from selected developing countries. J. Infect. Dev. Ctries 9: 1-7. https://doi.org/10.3855/jidc.5065
  38. Nurul Najian AB, Engku Nur Syafirah EA, Ismail N, Mohamed M, Yean CY. 2016. Development of multiplex loop mediated isothermal amplification (m-LAMP) labelbased gold nanoparticles lateral flow dipstick biosensor for detection of pathogenic Leptospira. Anal. Chim. Acta 903: 142-148. https://doi.org/10.1016/j.aca.2015.11.015
  39. Abdel-Aziz NM. 2016. Detection of Salmonella species in chicken carcasses using genus specific primer belong to invA gene in Sohag city, Egypt. Vet. World 9: 1125-1128. https://doi.org/10.14202/vetworld.2016.1125-1128
  40. Hoenigl M, Prattes J, Spiess B, Wagner J, Prueller F, Raggam RB, et al. 2014. Performance of galactomannan, beta-d-glucan, Aspergillus lateral-flow device, conventional culture, and PCR tests with bronchoalveolar lavage fluid for diagnosis of invasive pulmonary aspergillosis. J. Clin. Microbiol. 52: 2039-2045. https://doi.org/10.1128/JCM.00467-14
  41. Mashooq M, Kumar D, Niranjan AK, Agarwal RK, Rathore R. 2016. Development and evaluation of probe based real time loop mediated isothermal amplification for Salmonella: A new tool for DNA quantification. J. Microbiol. Methods 126: 24-29. https://doi.org/10.1016/j.mimet.2016.04.014
  42. Kokkinos PA, Ziros PG, Bellou M, Vantarakis A. 2014. Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Salmonella in Food. Food Analytical Methods 7: 512-526. https://doi.org/10.1007/s12161-013-9748-8
  43. Demeke T, Jenkins GR. 2010. Influence of DNA extraction methods, PCR inhibitors and quantification methods on real-time PCR assay of biotechnology-derived traits. Anal. Bioanal. Chem. 396: 1977-1990. https://doi.org/10.1007/s00216-009-3150-9
  44. Lu YP, Cheng J, Jiang SF, Zhang LW, Gao ZY, Han B, et al. 2010. Development of multiple quantitative fluorescent PCR for rapid diagnosis of common aneuploidy and it’s clinical application. Yi Chuan 32: 1141-1146.
  45. Wang C, Shen X, Lu J, Zhang L. 2013. Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system for rapid detection of HDV genotype 1. Lett. Appl. Microbiol. 56: 229-235. https://doi.org/10.1111/lam.12039
  46. Domesle KJ, Yang Q, Hammack TS, Ge B. 2018. Validation of a Salmonella loop-mediated isothermal amplification assay in animal food. Int. J. Food Microbiol. 264: 63-76. https://doi.org/10.1016/j.ijfoodmicro.2017.10.020
  47. Iseki H, Alhassan A, Ohta N, Thekisoe OM, Yokoyama N, Inoue N, et al. 2007. Development of a multiplex loopmediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites. J. Microbiol. Methods 71: 281-287. https://doi.org/10.1016/j.mimet.2007.09.019
  48. Wong YP, Othman S, Lau YL, Son R, Chee HY. 2017. Loop mediated isothermal amplification (LAMP): a versatile technique for detection of microorganisms. J. Appl. Microbiol. 300: 83-89.

Cited by

  1. Establishment and Application of Polymerase Spiral Reaction Amplification for Salmonella Detection in Food vol.29, pp.10, 2019, https://doi.org/10.4014/jmb.1906.06027
  2. Loop-mediated isothermal amplification (LAMP) - review and classification of methods for sequence-specific detection vol.12, pp.6, 2019, https://doi.org/10.1039/c9ay02246e
  3. Non-Invasive Colorimetric Magneto Loop-Mediated Isothermal Amplification (CM-LAMP) Method for Helicobacter pylori Detection vol.31, pp.4, 2019, https://doi.org/10.4014/jmb.2101.01008
  4. Rapid and simultaneous detection of Campylobacter spp. and Salmonella spp. in chicken samples by duplex loop-mediated isothermal amplification coupled with a lateral flow biosensor assay vol.16, pp.7, 2019, https://doi.org/10.1371/journal.pone.0254029
  5. Detection of harmful foodborne pathogens in food samples at the points of sale by MALDT-TOF MS in Egypt vol.14, pp.1, 2019, https://doi.org/10.1186/s13104-021-05533-8