Advertisement

Bioprocess and Biosystems Engineering

, Volume 41, Issue 5, pp 603–611 | Cite as

Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish

  • Weifang Gao
  • Hailong Huang
  • Peng Zhu
  • Xiaojun Yan
  • Jianzhong Fan
  • Jinpo Jiang
  • Jilin Xu
Research Paper
  • 238 Downloads

Abstract

Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex to detect contamination of bacteria in the food chain in a timely manner, and few studies have aimed to identify Salmonella in shellfish early in the supply chain. We herein developed a method for rapid detection of Salmonella in shellfish based on the method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD), which targets the invasion gene A (invA). The RPA-LFD was able to function at 30–45 °C, and at the temperature of 40 °C, it only took 8 min of amplification to reach the test threshold of amplicons. The established method had both a good specificity and a sensitivity of 100 fg DNA per reaction (20 µL). Regarding practical performance, RPA-LFD performed better than real-time PCR. Another advantage of RPA-LFD is that it was capable of being performed without expensive equipments. Thus, RPA-LFD has potential for further development as a detection kit for Salmonella in shellfish and other foods under field conditions.

Keywords

Recombinase polymerase amplification Lateral flow dipstick Nucleic acid test Salmonella Shellfish 

Notes

Acknowledgements

Authors would like to acknowledge the Ningbo Academy of Inspection and Quarantine for providing the Salmonella strain, DNAs of other serotypes of Salmonella and non-Salmonella organisms for this study, and their allowing us to finish assays of cultivating strains in the laboratory there. Authors also appreciated the assistance and guidance of the staff there. We thank LetPub (http://www.letpub.com) for their linguistic assistance during the preparation of this manuscript.

Funding

This study was funded by Ningbo Innovation Team (2015C110018), Ningbo Science and Technology Research Projects (2017C110003), Zhejiang Provincial Public Welfare Technology Program of China (2017C33133), K.C. Wang Magna Fund in Ningbo University (SS), Scientific Research Foundation of Graduate School of Ningbo University (G16091), the Earmarked Fund for Modern Agro-industry Technology Research System, China (CARS-49) and Zhejiang Xinmiao Talents Program (2015R405013).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Lund BM (2015) Microbiological food safety for vulnerable people. Int J Environ Res Public Health 12:10117–10132CrossRefGoogle Scholar
  2. 2.
    Botelho-Nevers E, Gautret P (2013) Outbreaks associated to large open air festivals, including music festivals, 1980 to 2012. Euro Surveill 18:20426CrossRefGoogle Scholar
  3. 3.
    Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, Jones JL, Griffin PM (2011) Foodborne illness acquired in the United States—major pathogens. Emerg Infect Dis 17:7–15CrossRefGoogle Scholar
  4. 4.
    EFSA ECDC. (2013) The European Union Summary Report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2011. EFSA J 11:3129CrossRefGoogle Scholar
  5. 5.
    Ranieri ML, Shi C, Switt AIM, Bakker HCD, Wiedmann M (2013) Comparison of typing methods with a new procedure based on sequence characterization for Salmonella serovar prediction. J Clin Microbiol 51:1786–1797CrossRefGoogle Scholar
  6. 6.
    Foley SL, Lynne AM (2008) Food animal-associated Salmonella challenges: pathogenicity and antimicrobial resistance. J Anim Sci 86:173–187CrossRefGoogle Scholar
  7. 7.
    Mąka Ł, Popowska M (2016) Antimicrobial resistance of Salmonella spp. isolated from food. Rocz Panstw Zakl Hig 67:343–358Google Scholar
  8. 8.
    Eady M, Park B (2016) Rapid identification of Salmonella serotypes through hyperspectral microscopy with different lighting sources. J Spectr Imaging 5:a4CrossRefGoogle Scholar
  9. 9.
    He Y, Lin L, Alam MJ, Shinoda S, Miyoshi S, Lei S (2010) Prevalence and antimicrobial resistance of Salmonella in retail foods in northern China. Int J Food Microbiol 143:230–234CrossRefGoogle Scholar
  10. 10.
    EFSA ECDC. (2015) The European Union Summary Report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2014. EFSA J 13:4329Google Scholar
  11. 11.
    OzFoodNet Working Group (2012) Monitoring the incidence and causes of diseases potentially transmitted by food in Australia: annual report of the OzFoodNet network, 2010. Commun Dis Intell Q Rep 36:E213–E241Google Scholar
  12. 12.
    Wu H, Xia X, Cui Y, Hu Y, Xi M, Wang X, Shi X, Wang D, Meng J, Yang B (2013) Prevalence of extended-spectrum beta-lactamase-producing Salmonella on; retail chicken in six provinces and two national cities in the People’s Republic of China. J Food Prot 76:2040–2044CrossRefGoogle Scholar
  13. 13.
    Xiong D, Song L, Geng S, Tao J, An S, Pan Z, Jiao X (2016) One-step PCR detection of Salmonella Pullorum/Gallinarum using a novel target: the flagellar biosynthesis gene flhB. Front Microbiol 7:1863Google Scholar
  14. 14.
    O’Regan E, Mccabe E, Burgess C, Mcguinness S, Barry T, Duffy G, Whyte P, Fanning S (2008) Development of a real-time multiplex PCR assay for the detection of multiple Salmonella serotypes in chicken samples. BMC Microbiol 8:156CrossRefGoogle Scholar
  15. 15.
    Wang L, Shi L, Alam MJ, Geng Y, Li L (2008) Specific and rapid detection of foodborne Salmonella by loop-mediated isothermal amplification method. Food Res Int 41:69–74CrossRefGoogle Scholar
  16. 16.
    Wang R, Ni Y, Xu Y, Jiang Y, Dong C, Na C (2015) Immuno-capture and in situ detection of Salmonella typhimurium on a novel microfluidic chip. Anal Chim Acta 853:710–717CrossRefGoogle Scholar
  17. 17.
    Pablos C, Marugán J, Cristóbal S, Grieken Rv (2017) Implications of electrical impedance-based microbiological technology in pork meat processing industry for the rapid detection and quantification of Salmonella Spp. J Food Sci Eng 7:1–16Google Scholar
  18. 18.
    Spector MP (1998) The starvation-stress response (SSR) of Salmonella. Adv Microb Physiol 40:233–279CrossRefGoogle Scholar
  19. 19.
    Kersting S, Rausch V, Bier FF, Nickisch-Rosenegk Mv (2014) Multiplex isothermal solid-phase recombinase polymerase amplification for the specific and fast DNA-based detection of three bacterial pathogens. Mikrochim Acta 181:1715–1723CrossRefGoogle Scholar
  20. 20.
    Rahn K, De Grandis SA, Clarke RC, Mcewen SA, Galán JE, Ginocchio C, Curtiss RI, Gyles CL (1992) Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Mol Cell Probes 6:271–279CrossRefGoogle Scholar
  21. 21.
    González-Escalona N, Brown EW, Zhang G (2012) Development and evaluation of a multiplex real-time PCR (qPCR) assay targeting ttrRSBCA locus and invA gene for accurate detection of Salmonella spp. in fresh produce and eggs. Food Res Int 48:202–208CrossRefGoogle Scholar
  22. 22.
    Malorny B, Hoorfar J, Bunge C, Helmuth R (2003) Multicenter validation of the analytical accuracy of Salmonella PCR: towards an international standard. Appl Environ Microbiol 69:290–296CrossRefGoogle Scholar
  23. 23.
    Jia Y, Mak PI, Massey C, Martins RP, Wangh LJ (2013) Construction of a microfluidic chip, using dried-down reagents, for LATE-PCR amplification and detection of single-stranded DNA. Lab Chip 13:4635–4641CrossRefGoogle Scholar
  24. 24.
    Gangwar M, Waters AM, Bej GA, Bej AK, Mojib N (2013) Detection of Salmonella in shellfish using SYBR Green I-based real-time multiplexed PCR assay targeting invA and spvB. Food Anal Methods 6:922–932CrossRefGoogle Scholar
  25. 25.
    Lazcka O, Del Campo FJ, Muñoz FX (2007) Pathogen detection: a perspective of traditional methods and biosensors. Biosens Bioelectron 22:1205–1217CrossRefGoogle Scholar
  26. 26.
    Piepenburg O, Williams CH, Stemple DL, Armes NA (2006) DNA detection using recombination proteins. PLoS Biol 4:1115–1121CrossRefGoogle Scholar
  27. 27.
    Jia L, Joanne M (2015) Advances in isothermal amplification: novel strategies inspired by biological processes. Biosens Bioelectron 64:196–211CrossRefGoogle Scholar
  28. 28.
    del Río JS, Adly NY, Acero-Sánchez JL, Henry OYF, O’Sullivan CK (2014) Electrochemical detection of Francisella tularensis genomic DNA using solid-phase recombinase polymerase amplification. Biosens Bioelectron 54:674–678CrossRefGoogle Scholar
  29. 29.
    Ahmed A, Linden van der H, Hartskeerl RA (2014) Development of a recombinase polymerase amplification assay for the detection of pathogenic Leptospira. Int J Environ Res Public Health 11:4953–4964CrossRefGoogle Scholar
  30. 30.
    Murinda SE, Ibekwe AM, Zulkaffly S, Cruz A, Park S, Razak N, Paudzai FM, Ab Samad L, Baquir K, Muthaiyah K, Santiago B, Rusli A, S B (2014) Real-time isothermal detection of Shiga toxin-producing Escherichia coli using recombinase polymerase amplification. Foodborne Pathog Dis 11:529–536CrossRefGoogle Scholar
  31. 31.
    Gao W, Huang H, Zhang Y, Zhu P, Yan X, Fan J, Chen X (2017) Recombinase polymerase amplification-based assay for rapid detection of listeria monocytogenes in food samples. Food Anal Methods 10:1972–1981CrossRefGoogle Scholar
  32. 32.
    Crannell ZA, Castellanosgonzales A, Nair G, Mejia R, White AC, Richardskortum R (2015) A multiplexed recombinase polymerase amplification assay to detect intestinal protozoa. Anal Chem 88:1610–1616CrossRefGoogle Scholar
  33. 33.
    Castellanos-Gonzalez A, Saldarriaga OA, Tartaglino L, Gacek R, Temple E, Sparks H, Melby PC, Travi BL (2015) A novel molecular test to diagnose canine visceral leishmaniasis at the point of care. Am J Trop Med Hyg 93:970–975CrossRefGoogle Scholar
  34. 34.
    Yang Y, Qin X, Wang G, Zhang Y, Shang Y, Zhang Z (2015) Development of a fluorescent probe-based recombinase polymerase amplification assay for rapid detection of Orf virus. Virol J 12:206CrossRefGoogle Scholar
  35. 35.
    Faye O, Faye O, Soropogui B, Patel P, El Wahed AA, Loucoubar C, Fall G, Kiory D, Magassouba N, Keita S, Kondé MK, Diallo AA, Koivogui L, Karlberg H, Mirazimi A, Nentwich O, Piepenburg O, Niedrig M, Weidmann M, Sall AA (2015) Development and deployment of a rapid recombinase polymerase amplification Ebola virus detection assay in Guinea in 2015. Euro Surveill 20:S.1–9Google Scholar
  36. 36.
    Liu W, Liu HX, Zhang L, Hou XX, Wan KL, Hao Q (2016) A novel isothermal assay of Borrelia burgdorferi by recombinase polymerase amplification with lateral flow detection. Int J Mol Sci 17:1250CrossRefGoogle Scholar
  37. 37.
    Prescott MA, Reed AN, Jin L, Pastey MK (2016) Rapid detection of cyprinid herpesvirus 3 in latently infected Koi by recombinase polymerase amplification. J Aquat Anim Health 28:173–180CrossRefGoogle Scholar
  38. 38.
    Sun K, Xing W, Yu X, Fu W, Wang Y, Zou M, Luo Z, Xu D (2016) Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of Schistosoma japonicum. Parasites Vectors 9:476CrossRefGoogle Scholar
  39. 39.
    Lillis L, Siverson J, Lee A, Cantera J, Parker M, Piepenburg O, Lehman DA, Boyle DS (2016) Factors influencing recombinase polymerase amplification (RPA) assay outcomes at point of care. Mol Cell Probes 30:74–78CrossRefGoogle Scholar
  40. 40.
    Boyle DS, McNerney R, Low HT, Leader BT, Pe´rez-Osorio AC, Meyer JC, O’Sullivan DM, Brooks DG, Piepenburg O, Forrest MS (2014) Rapid detection of mycobacterium tuberculosis by recombinase polymerase amplification. PLoS One 9:e103091CrossRefGoogle Scholar
  41. 41.
    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 62:20–29CrossRefGoogle Scholar
  42. 42.
    Kersting S, Rausch V, Bier FF, Nickisch-Rosenegk Mv (2014) Rapid detection of plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis. Malar J 13:99CrossRefGoogle Scholar
  43. 43.
    Han F, Ge B (2008) Evaluation of a loop-mediated isothermal amplification assay for detecting Vibrio vulnificus in raw oysters. Foodborne Pathog Dis 5:311CrossRefGoogle Scholar
  44. 44.
    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 11:574–580CrossRefGoogle Scholar
  45. 45.
    Guo H (2010) Study on rapid detection method by multiple PCR of four species animal food borne pathogenic bacteria. Master’s thesis, Heilongjiang Bayi Agricultural University, Heilongjiang ProvinceGoogle Scholar
  46. 46.
    Zhang W, Xie Z, Zuo H, Ding X, Pei X (2010) Detection of food-borne pathogens with polymerase chain reaction and introduction of food safety supervision system in China. Qual Assur Saf Crops Foods 2:13–21CrossRefGoogle Scholar
  47. 47.
    Lillis L, Lehman D, Singhal MC, Cantera J, Singleton J, Labarre P, Toyama A, Piepenburg O, Parker M, Wood R, Overbaugh J, Boyle DS (2014) Non-instrumented incubation of a recombinase polymerase amplification assay for the rapid and sensitive detection of proviral HIV-1 DNA. PLoS One 9:e108189CrossRefGoogle Scholar
  48. 48.
    Ahn YC, Cho MH, Yoon IK, Jung DH, Lee EY, Kim JH, Jang WC, Ahn YC, Cho MH, Yoon IK (2010) Detection of Salmonella using the loop mediated isothermal amplification and real-time PCR. J Korean Chem Soc 54:215–221CrossRefGoogle Scholar
  49. 49.
    Zhuang L, Gong J, Li Q, Zhu C, Yu Y, Dou X, Liu X, Xu B, Wang C (2014) Detection of Salmonella spp. by a loop-mediated isothermal amplification (LAMP) method targeting bcfD gene. Lett Appl Microbiol 59:658–664CrossRefGoogle Scholar
  50. 50.
    Hiroshi K, Jun K, Kumiko F, Kenichi H, Masayoshi I, Naomi F, Mika N, Shunichi O, Toshiya S, Kazushige T (2009) Simultaneous enrichment of Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes by single broth and screening of the pathogens by multiplex real-time PCR. Food Sci Technol Res 15:427–438Google Scholar
  51. 51.
    Kim TH, Park J, Kim CJ, Cho YK (2014) Fully integrated lab-on-a-disc for nucleic acid analysis of food-borne pathogens. Anal Chem 86:3841–3848CrossRefGoogle Scholar
  52. 52.
    Ji YK, Lee JL (2016) Rapid detection of Salmonella enterica serovar enteritidis from eggs and chicken meat by real-time recombinase polymerase amplification in comparison with the two-step real-time PCR. J Food Saf 32:611–615Google Scholar
  53. 53.
    Choi G, Jung JH, Park BH, Oh SJ, Seo JH, Choi JS, Kim H, Seo TS (2016) A centrifugal direct recombinase polymerase amplification (direct-RPA) microdevice for multiplex and real-time identification of food poisoning bacteria. Lab Chip 16:2309–2316CrossRefGoogle Scholar
  54. 54.
    Katrin K, Jekaterina F, Oana T, Julia S, Taavi L, Hiljar S, Imre M, Made L, Indrek T, Ülo L (2014) Sensitive and rapid detection of Chlamydia trachomatis by recombinase polymerase amplification directly from urine samples. J Mol Diagn JMD 16:127–135CrossRefGoogle Scholar
  55. 55.
    Valiadi M, Kalsi S, Jones IGF, Turner C, Sutton JM, Morgan H (2016) Simple and rapid sample preparation system for the molecular detection of antibiotic resistant pathogens in human urine. Biomed Microdevices 18:18CrossRefGoogle Scholar
  56. 56.
    Rosser A, Rollinson D, Forrest M, Webster BL (2015) Isothermal recombinase polymerase amplification (RPA) of Schistosoma haematobium DNA and oligochromatographic lateral flow detection. Parasites Vectors 8:446CrossRefGoogle Scholar
  57. 57.
    Liljander A, Yu M, O’Brien E, Heller M, Nepper JF, Weibel DB, Gluecks I, Younan M, Frey J, Falquet L (2015) Field-applicable recombinase polymerase amplification assay for rapid detection of Mycoplasma capricolum subsp. capripneumoniae. J Clin Microbiol 53:2810–2815CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Weifang Gao
    • 1
    • 2
  • Hailong Huang
    • 1
  • Peng Zhu
    • 1
    • 2
  • Xiaojun Yan
    • 1
    • 2
  • Jianzhong Fan
    • 3
  • Jinpo Jiang
    • 4
  • Jilin Xu
    • 1
  1. 1.Ningbo UniversityNingboChina
  2. 2.Ningbo Institute of OceanographyNingboChina
  3. 3.Ningbo Boao Biological Engineering Co., Ltd.NingboChina
  4. 4.Ningbo City College of Vocational TechnologyNingboChina

Personalised recommendations