Skip to main content

Advertisement

SpringerLink
Log in

High-flux simultaneous screening of common foodborne pathogens and their virulent factors

  • Research Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4–7 days to complete. In this study, we described a high-flux polymerase chain reaction (PCR) method for simultaneous detection of nine targeted genes (rfbE, stx1, stx2, invA, oprI, tlh, trh, tdh, and hlyA) with multiplex strains. The designed primers were highly specific for their respective target gene fragments. As the selected primers follow the principles of similar melting and annealing temperature, all the targeted genes could be detected for one strain with the same PCR program. Combining with 96-well PCR plate, by adding a single different gene to each well in each row, both the ATCC strains (E. coli, Salmonella spp., V. parahaemolyticus, L. monocytogenes, P. aeruginosa, S. aureus) and the clinical strains (E. coli, P. aeruginosa, S. aureus) were simultaneously detected to carry their specific and virulence genes. Therefore, using 96-well PCR plate for PCR amplification might be applied to high-flux sequencing of specific and virulence genes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Mcentire J (2013) Foodborne disease: the global movement of food and people. Infect Dis Clin N Am 27:687–693

    Article  Google Scholar 

  2. Chen DQ, Yang L, Peters BM et al (2019) Complete sequence of a novel multidrug-resistant pseudomonas putida strain carrying two copies of qnrVC6. Microb Drug Resist 25:1–7

    Article  PubMed  CAS  Google Scholar 

  3. Zhao XH, Yu ZX, Xu ZB (2018) Study the features of 57 confirmed CRISPR Loci in 38 strains of Staphylococcus aureus. Front Microbiol 9:1591

    Article  PubMed  PubMed Central  Google Scholar 

  4. Wen SX, Feng DH, Lu ZR et al (2018) Microbial infection pattern, pathogenic features and resistance mechanism of carbapenem-resistant Gram negative bacilli during long-term hospitalization. Microb Pathogenesis 117:356–360

    Article  CAS  Google Scholar 

  5. Wu Q, Ye Y, Li F et al (2016) Prevalence and genetic characterization of Pseudomonas aeruginosa in drinking water in Guangdong Province of China. LWT Food Sci Technol 69:24–31

    Article  CAS  Google Scholar 

  6. Wang F, Jiang L, Yang Q et al (2011) Prevalence and antimicrobial susceptibility of major foodborne pathogens in imported seafood. J Food Prot 74:1451–1461

    Article  PubMed  Google Scholar 

  7. Chao G, Zhou X, Jiao X et al (2007) Prevalence and antimicrobial resistance of foodborne pathogens isolated from food products in China. Foodborne Pathog Dis 4:277–284

    Article  PubMed  CAS  Google Scholar 

  8. Newell DG, Koopmans M, Verhoef L et al (2010) Food-borne diseases—the challenges of 20 years ago still persist while new ones continue to emerge. Int J Food Microbiol 139(Suppl 1):S3–15

    Article  PubMed  Google Scholar 

  9. Liu JY, Yang L, Hou YC et al (2018) Transcriptomics study on Staphylococcus aureus biofilm under low concentration of ampicillin. Front Microbiol 9:2413

    Article  PubMed  PubMed Central  Google Scholar 

  10. Miao J, Liang YR, Chen LQ et al (2017) Formation and development of Staphylococcus biofilm: with focus on food safety. J Food Saf 37:e12358

    Article  Google Scholar 

  11. Bao XR, Jia XY, Chen LQ et al (2017) Effect of polymyxin resistance (pmr) on biofilm formation of Cronobacter sakazakii. Microb Pathogenesis 106:16–19

    Article  CAS  Google Scholar 

  12. Xu ZB, Liang YR, Lin SQ et al (2016) Crystal violet and XTT assays on Staphylococcus aureus biofilm quantification. Curr Microbiol 73:474–482

    Article  PubMed  CAS  Google Scholar 

  13. Xue J, Zhang W (2013) Understanding China's food safety problem: an analysis of 2387 incidents of acute foodborne illness. Food Control 30:311–317

    Article  Google Scholar 

  14. Liu LY, Xu RR, Li L et al (2018) Correlation and in vitro mechanism of bactericidal activity on E. coli with whey protein isolate during ultrasonic treatment. Microb Pathogenesis 115:154–158

    Article  CAS  Google Scholar 

  15. Liu LY, Lu ZR, Li L et al (2018) Physical relation and mechanism of ultrasonic bactericidal activity on pathogenic E. coli with WPI. Microb Pathogenesis 117:73–79

    Article  Google Scholar 

  16. Liu JY, Li L, Zhou LZ et al (2018) Effect of ultrasonic field on the enzyme activities and ion balance of potential pathogen Saccharomyces cerevisiae. Microb Pathogenesis 119:216–220

    Article  CAS  Google Scholar 

  17. Jia XY, Hua JJ, Liu L et al (2018) Phenotypic characterization of pathogenic Cronobacter spp. strains. Microb Pathogenesis 121:232–237

    Article  CAS  Google Scholar 

  18. Xu ZB, Xu XY, Qi D et al (2017) Effect of aminoglycosides on the pathogenic characteristics of microbiology. Microb Pathogenesis 113:357–364

    Article  CAS  Google Scholar 

  19. Xu ZB, Hou YC, Peters BM et al (2016) Chromogenic media for MRSA diagnostics. Mol Biol Rep 43:1205–1212

    Article  PubMed  CAS  Google Scholar 

  20. Zhao X, Lin CW, Wang J et al (2014) Advances in rapid detection methods for foodborne pathogens. J Microbiol Biotechnol 24:297–312

    Article  PubMed  CAS  Google Scholar 

  21. Zhao XH, Li M, Xu ZB (2018) Detection of foodborne pathogens by surface enhanced Raman spectroscopy. Front Microbiol 9:1236

    Article  PubMed  PubMed Central  Google Scholar 

  22. Miao J, Wang WX, Xu WY et al (2018) The fingerprint mapping and genotyping systems application on methicillin-resistant Staphylococcus aureus. Microb Pathogenesis 125:246–251

    Article  CAS  Google Scholar 

  23. Miao J, Chen LQ, Wang JW et al (2017) Current methodologies on genotyping for nosocomial pathogen methicillin-resistant Staphylococcus aureus (MRSA). Microb Pathogenesis 107:17–28

    Article  CAS  Google Scholar 

  24. Bao XR, Yang L, Chen LQ et al (2017) Analysis on pathogenic and virulent characteristics of the Cronobacter sakazakii strain BAA-894 by whole genome sequencing and its demonstration in basic biology science. Microb Pathogenesis 109:280–286

    Article  CAS  Google Scholar 

  25. Bao XR, Yang L, Chen LQ et al (2017) Virulent and pathogenic features on the Cronobacter sakazakii polymyxin resistant pmr mutant strain s-3. Microb Pathogenesis 110:359–364

    Article  CAS  Google Scholar 

  26. Lin SQ, Yang L, Chen G et al (2017) Pathogenic features and characteristics of food borne pathogens biofilm: biomass, viability and matrix. Microb Pathogenesis 111:285–291

    Article  CAS  Google Scholar 

  27. Xu ZB, Xie JH, Liu JY et al (2017) Whole-genome resequencing of Bacillus cereus and expression of genes functioning in sodium chloride stress. Microb Pathogenesis 104:248–253

    Article  CAS  Google Scholar 

  28. Yu GC, Wen WR, Peters BM et al (2016) First report of novel genetic array aacA4-bla(IMP-25)-oxa30-catB3 and identification of novel metallo-beta-lactamase gene bla(IMP25): a Retrospective Study of antibiotic resistance surveillance on Psuedomonas aeruginosa in Guangzhou of South China, 2003–2007. Microb Pathogenesis 95:62–67

    Article  CAS  Google Scholar 

  29. Xu ZB, Xie JH, Yang L et al (2018) Complete sequence of pCY-CTX, a plasmid carrying a phage-like region and an ISEcp1-mediated Tn2 element from Enterobacter cloacae. Microb Drug Resist 24:307–313

    Article  PubMed  CAS  Google Scholar 

  30. Desmarchelier PM, Bilge SS, Fegan N et al (1998) A PCR specific for Escherichia coli O157 based on the rfb locus encoding O157 lipopolysaccharide. J Clin Microbiol 36:1801–1804

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Rahn K, De Grandis SA, Clarke RC et al (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 6:271–279

    Article  PubMed  CAS  Google Scholar 

  32. Matthijs S, Coorevits A, Gebrekidan TT et al (2013) Evaluation of oprI and oprL genes as molecular markers for the genus Pseudomonas and their use in studying the biodiversity of a small Belgian River. Res Microbiol 164:254–261

    Article  PubMed  CAS  Google Scholar 

  33. Rawool DB, Malik SV, Shakuntala I et al (2007) Detection of multiple virulence-associated genes in Listeria monocytogenes isolated from bovine mastitis cases. Int J Food Microbiol 113:201–207

    Article  PubMed  CAS  Google Scholar 

  34. Bej AK, Patterson DP, Brasher CW et al (1999) Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. J Microbiol Meth 36:215–225

    Article  CAS  Google Scholar 

  35. Xu ZB, Li L, Shi L et al (2011) Class 1 integron in staphylococci. Mol Biol Rep 38:5261–5279

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. Xu Z, Li L, Shirtliff ME et al (2011) Resistance class 1 integron in clinical methicillin-resistant Staphylococcus aureus strains in southern China, 2001–2006. Clin Microbiol Infec 17:714–718

    Article  CAS  Google Scholar 

  37. Xu ZB, Li L, Shirtliff ME et al (2010) First report of class 2 integron in clinical Enterococcus faecalis and class 1 integron in Enterococcus faecium in South China. Diagn Micr Infect Dis 68:315–317

    Article  CAS  Google Scholar 

  38. Xu ZB, Li L, Shirtliff ME et al (2009) Occurrence and characteristics of Class 1 and 2 integrons in Pseudomonas aeruginosa Isolates from patients in Southern China. J Clin Microbiol 47:230–234

    Article  PubMed  CAS  Google Scholar 

  39. Xu ZB, Shi L, Alam MJ et al (2008) Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004. Fems Microbiol Lett 278:223–230

    Article  PubMed  CAS  Google Scholar 

  40. Xu ZB, Li L, Alam MJ et al (2008) First confirmation of integron-bearing methicillin-resistant Staphylococcus aureus. Curr Microbiol 57:264–268

    Article  PubMed  CAS  Google Scholar 

  41. Xu Z, Shi L, Zhang C et al (2007) Nosocomial infection caused by class 1 integron-carrying Staphylococcus aureus in a hospital in South China. Clin Microbiol Infec 13:980–984

    Article  CAS  Google Scholar 

  42. Deng Y, Liu JY, Peters BM et al (2015) Antimicrobial resistance investigation on Staphylococcus strains in a local hospital in Guangzhou, China, 2001–2010. Microb Drug Resist 21:102–104

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Liu JY, Chen DQ, Peters BM et al (2016) Staphylococcal chromosomal cassettes mec (SCCmec): a mobile genetic element in methicillin-resistant Staphylococcus aureus. Microb Pathogenesis 101:56–67

    Article  CAS  Google Scholar 

  44. Miao J, Peters BM, Li L et al (2016) Evaluation of Eric-Pcr for fingerprinting methicillin-resistant Staphylococcus aureus Strains. Basic Clin Pharmacol 118:33–33

    Google Scholar 

  45. Miao J, Chen LQ, Wang JW et al (2017) Evaluation and application of molecular genotyping on nosocomial pathogen-methicillin-resistant Staphylococcus aureus isolates in Guangzhou representative of Southern China. Microb Pathogenesis 107:397–403

    Article  CAS  Google Scholar 

  46. Xu ZB, Xie JH, Peters BM et al (2017) Longitudinal surveillance on antibiogram of important Gram-positive pathogens in Southern China, 2001–2015. Microb Pathogenesis 103:80–86

    Article  CAS  Google Scholar 

  47. Xie JH, Yang L, Peters BM et al (2017) A 16-year retrospective surveillance report on the pathogenic features and antimicrobial susceptibility of Pseudomonas aeruginosa isolates from FAHJU in Guangzhou representative of Southern China. Microb Pathogenesis 110:37–41

    Article  CAS  Google Scholar 

  48. Xie JH, Peters BM, Li B et al (2017) Clinical features and antimicrobial resistance profiles of important Enterobacteriaceae pathogens in Guangzhou representative of Southern China, 2001–2015. Microb Pathogenesis 107:206–211

    Article  CAS  Google Scholar 

  49. Xu ZB, Li L, Chu J et al (2012) Development and application of loop-mediated isothermal amplification assays on rapid detection of various types of Staphylococci strains. Food Res Int 47:166–173

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Wang L, Li Y, Chu J et al (2012) Development and application of a simple loop-mediated isothermal amplification method on rapid detection of Listeria monocytogenes strains. Mol Biol Rep 39:445–449

    Article  PubMed  CAS  Google Scholar 

  51. Zhao XH, Wang L, Li YM et al (2011) Development and application of a loop-mediated isothermal amplification method on rapid detection of Pseudomonas aeruginosa strains. World J Microb Biot 27:181–184

    Article  CAS  Google Scholar 

  52. Xu ZB, Li L, Zhao XH et al (2011) Development and application of a novel multiplex polymerase chain reaction (PCR) assay for rapid detection of various types of staphylococci strains. Afr J Microbiol Res 5:1869–1873

    CAS  Google Scholar 

  53. Wang L, Zhao XH, Chu J et al (2011) Application of an improved loop-mediated isothermal amplification detection of Vibrio parahaemolyticus from various seafood samples. Afr J Microbiol Res 5:5765–5771

    Article  CAS  Google Scholar 

  54. Zhao XH, Wang L, Chu J et al (2010) Rapid detection of vibrio parahaemolyticus strains and virulent factors by loop-mediated isothermal amplification assays. Food Sci Biotechnol 19:1191–1197

    Article  Google Scholar 

  55. Zhao XH, Wang L, Chu J et al (2010) Development and application of a rapid and simple loop-mediated isothermal amplification method for food-borne salmonella detection. Food Sci Biotechnol 19:1655–1659

    Article  Google Scholar 

  56. Zhao XH, Li YM, Wang L et al (2010) Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples. Mol Biol Rep 37:2183–2188

    Article  PubMed  CAS  Google Scholar 

  57. Xu ZB, Hou YC, Qin D et al (2016) Evaluation of current methodologies for rapid identification of methicillin-resistant Staphylococcus aureus strains. Basic Clin Pharmacol 118:33–33

    Google Scholar 

  58. Ferrario C, Lugli GA, Ossiprandi MC et al (2017) Next generation sequencing-based multigene panel for high throughput detection of food-borne pathogens. Int J Food Microbiol 256:20–29

    Article  PubMed  CAS  Google Scholar 

  59. Liu JY, Zhou R, Li L et al (2017) Viable but non-culturable state and toxin gene expression of enterohemorrhagic Escherichia coli 0157 under cryopreservation. Res Microbiol 168:188–193

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Key R&D Program of China (no. 2016YFD0400203), YangFan Innovative and Entrepreneurial Research Team Project (No. 2014YT02S029), the National Key Research and Development Program of China (2016YFD04012021), Guangdong Special Support Program (2016TQ03N682), Pearl River S&T Nova Program of Guangzhou (201710010061), the Fundamental Research Funds for the Central Universities (Dr. Zhenbo Xu, D2191310), Collaborative Grant with AEIC (KEO-2019-0624-001-1), the 111 Project (B17018), and Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety (KL-2018-05, KL-2018-14, KL-2018-16).

Author information

Author notes

  1. Yanmei Li and Yisen Qiu contribute equally to this work.

Authors and Affiliations

  1. Department of Haematology, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, 510623, China

    Yanmei Li

  2. Department of Spine Surgery, The Second Affiliated Hospital of Shantou University Medical College, North Dongxia Road, Shantou, 515041, Guangdong, China

    Yisen Qiu

  3. Department of Dermato-Venereology, Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510640, China

    Congxiu Ye

  4. School of Food Science and Engineering, Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, South China University of Technology, Guangzhou, 510640, China

    Ling Chen & Junyan Liu

  5. Guangdong Zhongqing Font Biochemical Science and Technology Co. Ltd, Maoming, 525427, Guangdong, China

    Yi Liang

  6. Guangzhou KEO Biotechnology Co. LTD, Guangzhou, Guangdong, China

    Guoxing Liu

Authors
  1. Yanmei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yisen Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Congxiu Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ling Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yi Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Guoxing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Junyan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Guoxing Liu or Junyan Liu.

Ethics declarations

Conflict of interest

Author Yi Liang was employed by the company Guangdong Zhongqing Font Biochemical Science and Technology Co. Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, Y., Qiu, Y., Ye, C. et al. High-flux simultaneous screening of common foodborne pathogens and their virulent factors . Bioprocess Biosyst Eng 43, 693–700 (2020). https://doi.org/10.1007/s00449-019-02267-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-019-02267-7

Keywords

Search

Navigation