Skip to main content
SpringerLink
Log in

A fully integrated microdevice for capturing, amplification, and colorimetric detection of foodborne pathogens

  • Technical Paper
  • Published:
Microsystem Technologies Aims and scope Submit manuscript

Abstract

In the present study, we integrated laboratory functionalities of sample preparation, nucleic acid amplification, and colorimetric detection into a microdevice for the facile foodborne pathogen detection from bulk liquid samples. The developed microdevice consists of micropillars engraved in a microchamber. In order to capture pathogens from the high throughput sample, boronic acid was decorated on the surface of micropillars to exploit the high affinity of this functional group toward cis-diols of moieties constructing the bacterial outer membrane. Using disposable syringes, the surface modification process for the dense distribution of the boronic acid group was readily achieved at room temperature within 3 h. The success of surface modification was verified by the color changes and the fluorescence signals of the remnant alizarin red sodium. Escherichia coli O157:H7 involving green fluorescence protein (E. coli O157:H7 GFP) was used to evaluate the capture capability of the device at three inoculum bacterial concentrations. The temperature in the reaction zone was carefully investigated to ensure appropriate thermal conditions for performing chamber-type polymerase chain reaction. According to the results, the 584-bp sequence of the Shiga toxin gene of the pathogen was successfully amplified within 2 h. The amplicon was then visualized by the brown color of NaBH4-induced silver nanoparticles. Based on the results, it can be concluded that the proposed microdevice is a promising platform for robust detection of the foodborne pathogens in bulk liquid samples and is applicable in food industry, public healthcare, and environmental monitoring.

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
Fig. 4
Fig. 5

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

References

  • Agnihotri S, Mukherji S, Mukherji S (2014) Size-controlled silver nanoparticles synthesized over the range 5–100 nm using the same protocol and their antibacterial efficacy. RSC Adv 4:3974–3983

    Article  Google Scholar 

  • Ahrberg CD, Manz A, Chung BG (2016) Polymerase chain reaction in microfluidic devices. Lab Chip 16:3866–3884

    Article  Google Scholar 

  • Ali U, Karim KJBA, Buang NA (2015) A review of the properties and applications of poly(methyl methacrylate) (PMMA). Polym Rev 55:678–705

    Article  Google Scholar 

  • António JPM, Russo R, Carvalho CP, Cal PMSD, Gois PMP (2019) Boronic acids as building blocks for the construction of therapeutically useful bioconjugates. Chem Soc Rev 48:3513–3536

    Article  Google Scholar 

  • Berti L, Burley GA (2018) Nucleic acid and nucleotide-mediated synthesis of inorganic nanoparticles. Nature Nanotech 3:81–87

    Article  Google Scholar 

  • Chu YW, Engebretson DA, Carey JR (2013) Bioconjugated magnetic nanoparticles for the detection of bacteria. J Biomed Nanotech 9:1951–1961

    Article  Google Scholar 

  • Golabi M, Kuralay F, Jager EWH, Beni V, Turner APF (2017) Electrochemical bacterial detection using poly(3-aminophenylboronic acid)-based imprinted polymer. Biosens Bioelectron 93:87–93

    Article  Google Scholar 

  • Hall DG (2019) Boronic acid catalysis. Chem Soc Rev 48:3475–3496

    Article  Google Scholar 

  • Jung YL, Jung C, Park JH, Kim MI, Park HG (2013) Direct detection of unamplified genomic DNA based on photo-induced silver ion reduction by DNA molecules. Chem Commun 49:2350–2352

    Article  Google Scholar 

  • Kant K, Shahbazi MA, Dave VP, Ngo TA, Chidambara VA, Than LQ, Bang DD, Wolff A (2018) Microfluidic devices for sample preparation and rapid detection of foodborne pathogens. Biotech Advances 36:1003–1024

    Article  Google Scholar 

  • Kell J, Stewart G, Ryan S, Peytavi R, Boissinot M, Huletsky A, Bergeron MG, Simard B (2008) Vancomycin-modified nanoparticles for efficient targeting and preconcentration of gram-positive and gram-negative bacteria. ACS Nano 2:1777–1788

    Article  Google Scholar 

  • Kitagawa F, Kubota K, Sueyoshi K, Otsuka K (2006) One-step immobilization of cationic polymer onto a poly(methyl methacrylate) microchip for high-performance electrophoretic analysis of proteins. Sci Tech Ad Mater 7:558–565

    Article  Google Scholar 

  • Kuss S, Couto RAS, Evans RM, Lavender H, Tang CC, Compton RG (2019) Versatile electrochemical sensing platform for bacteria. Anal Chem 91:4317–4322

    Article  Google Scholar 

  • La HC, Lee NY (2019) Fabrication of a polycarbonate microdevice and boronic acid-mediated surface modification for on-chip sample purification and amplification of foodborne pathogens. Biomed Microdevice 21:72

    Article  Google Scholar 

  • Lacina K, Skládal P, James TD (2014) Boronic acids for sensing and other applications—a mini-review of papers published in 2013. Chem Cent J 8:60

    Article  Google Scholar 

  • Law JWF, Mutalib NSA, Chan KG, Lee LH (2014) Rapid methods for the detection of foodborne bacterial pathogens: principles, applications, advantages and limitations. Front Microbiol 5:770

    Google Scholar 

  • Li B, Tan H, Anastasova S, Power M, Seichepine F, Yang GZ (2019) A bio-inspired 3D micro-structure for graphene-based bacteria sensing. Biosens Bioelectron 123:77–84

    Article  Google Scholar 

  • Mairhofer J, Roppert K, Ertl P (2009) Microfluidic systems for pathogen sensing: a review. Sensors 9(6):4804–4823

    Article  Google Scholar 

  • Mavani K, Shah M (2013) Synthesis of silver nanoparticles by using sodium borohydride as a reducing agent. Int J Eng Res Technol 2(3):1–5

    Google Scholar 

  • Mei L, Busscher HJ, Mei HC, Ren Y (2011) Influence of surface roughness on streptococcal adhesion forces to composite resins. Dent Mater 27:770–778

    Article  Google Scholar 

  • Ocola LE, Costales M, Gosztola DJ (2016) Development characteristics of polymethyl methacrylate in alcohol/water mixtures: a lithography and Raman spectroscopy study. Nanotechnology 27:035302

    Article  Google Scholar 

  • Pearson B, Wang P, Mills A, Pang S, McLandsboroug L, He L (2017) Innovative sandwich assay with dual optical and SERS sensing mechanisms for bacterial detection. Anal Methods 9:4732–4739

    Article  Google Scholar 

  • Preedy E, Perni S, Nipic D, Bohinc K, Prokopovich P (2014) Surface roughness mediated adhesion forces between borosilicate glass and gram-positive bacteria. Langmuir 30:9466–9476

    Article  Google Scholar 

  • Ravindranath SP, Mauer LJ, Deb-Roy C, Irudayaraj J (2009) Biofunctionalized magnetic nanoparticle integrated mid-infrared pathogen sensor for food matrixes. Anal Chem 81(8):2840–2846

    Article  Google Scholar 

  • Rotem S, Raz N, Kashi Y, Mor A (2010) Bacterial capture by peptide-mimetic oligoacyllysine surfaces. Appl Environ Microbiol 76:3301–3307

    Article  Google Scholar 

  • Ryan S, Kell AJ, van Faassen H, Tay LL, Simard B, MacKenzie R, Gilbert M, Tanha J (2009) Single-domain antibody-nanoparticles: promising architectures for increased Staphylococcus aureus detection specificity and sensitivity. Bioconjugate Chem 20:1966–1974

    Article  Google Scholar 

  • Schrader C, Schielke A, Ellerbroek L, Johne R (2012) PCR inhibitors—occurrence, properties and removal. L Appl Microbiol 113:1014–1026

    Article  Google Scholar 

  • Shabbeer H, Khan A, Shah A, Rehman ZU, Shah SM, Khan A, Shah SS (2012) Effect of acidic and basic conditions on the plasmon band of colloidal silver. Walailak J Sci and Tech 9:229–237

    Google Scholar 

  • Sharma S, Jaimes-Lizcano YA, McLay RB, Cirino PC, Conrad JC (2016) Subnanometric roughness affects the deposition and mobile adhesion of Escherichia coli on silanized glass surfaces. Langmuir 32:5422–5433

    Article  Google Scholar 

  • Song Y, Lin B, Tian T, Xu X, Wang W, Ruan Q, Guo J, Zhu Z, Yang C (2019) Recent progress in microfluidics-based biosensing. Anal Chem 91:388–404

    Article  Google Scholar 

  • Springsteen G, Wang B (2002) Alizarin Red S as a general optical reporter for studying the binding of boronic acids with carbohydrates. Tetrahedron 58:5291–5300

    Article  Google Scholar 

  • Suresh SS, Mohanty S, Nayak SK (2017) Investigation into the mechanical and thermal properties of poly(methyl methacrylate) recovered from light guidance panels with a focus on future remanufacturing and sustainable waste management. J Remanufact 7:217–233

    Article  Google Scholar 

  • Trinh KTL, Chon NM, Lee NY (2018) Fabrication of an integrated polystyrene microdevice for pre-concentration and amplification of Escherichia coli O157:H7 from raw milk. Anal Methods 10:5071–5077

    Article  Google Scholar 

  • Trinh TND, La HC, Lee NY (2019) Fully integrated and foldable microdevice encapsulated with agarose for long-term storage potential for point-of-care testing of multiplex foodborne pathogens. ACS Sens 4:2754–2762

    Article  Google Scholar 

  • Truong VK, Lapovok R, Estrin YS, Rundell S, Wang J, Fluke CJ, Crawford RJ, Ivanova EP (2010) The influence of nano-scale surface roughness on bacterial adhesion to ultrafine-grained titanium. Biomaterials 31:3674–3683

    Article  Google Scholar 

  • Wang J, Gao J, Liu D, Han D, Wang Z (2012) Phenylboronic acid functionalized gold nanoparticles for highly sensitive detection of Staphylococcus aureus. Nanoscale 4:451–454

    Article  Google Scholar 

  • Wang P, Pang S, Pearson B, Chujo Y, McLandsborough L, Fan M, Lili H (2017) Rapid concentration detection and differentiation of bacteria in skimmed milk using surface enhanced Raman scattering mapping on 4-mercaptophenylboronic acid functionalized silver dendrites. Anal Bioanal Chem 409:2229–2238

    Article  Google Scholar 

  • Whyte GF, Vilar R, Woscholski R (2013) Molecular recognition with boronic acids-applications in chemical biology. Chem Biol 6:161–174

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. NRF-2017R1A2B4008179). This research was also supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI18C2096).

Author information

Authors and Affiliations

  1. Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam, Gyeonggi-do, 13120, Korea

    Hoang Chau La & Nae Yoon Lee

Authors
  1. Hoang Chau La
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nae Yoon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nae Yoon Lee.

Additional information

Publisher's Note

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

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (MP4 31966 kb)

Supplementary material 2 (DOCX 16 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

La, H.C., Lee, N.Y. A fully integrated microdevice for capturing, amplification, and colorimetric detection of foodborne pathogens. Microsyst Technol 26, 3875–3883 (2020). https://doi.org/10.1007/s00542-020-04877-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00542-020-04877-8

Search

Navigation