Chemical Papers

, Volume 74, Issue 2, pp 451–458 | Cite as

QCM immunosensor for the determination of Staphylococcus aureus antigen

  • Miroslav PohankaEmail author
Original Paper


Staphylococcus aureus is a causative agent of infectious diseases and a producer of a group of highly toxic proteins called Staphylococcal enterotoxins. Currently, the presence of S. aureus in a sample can be determined by a wide spectrum of genetic, immunochemical and microbiological techniques. Despite availability of standard methods and devices, they are not fully suitable for a simple, label-free mode of use by an unskilled worker. In this paper, a biosensor based on quartz crystal microbalance (QCM) sensor is performed as a tool for a simple and reliable determination of S. aureus. The biosensor contained an antibody specific to S. aureus immobilized through protein A on 10 MHz QCM sensors and the interdigitated layer was stabilized by iron nanoparticles. The assay was performed as label free and sample was applied without any pretreatment. Limit of detection 145 µg/ml (5.18 × 108 CFU/ml) was achieved. When considered standard size of a sample 50 μl, the limit of detection was equal to 7.25 μg (2.59 × 107 CFU). The biosensors kept more than 90% sensitivity to the analyte after storage in laboratory conditions for 90 days. Good correlation to standard enzyme-linked immunosorbent assay was also proved. The biosensor appears as a simple but reliable tool for S. aureus assay and the QCM platform would provide further applications in microbial analyses.


Affinity Antibody Biological weapon Immunosensor Label-free assay Piezoelectric Quartz crystal microbalance Staphylococcal enterotoxin Staphylococcus aureus 



This work was supported by a Ministry of Defence of the Czech Republic—long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence. Ms. Jitka Zakova is gratefully acknowledged for laboratory assistance.

Compliance with ethical standards

Conflicts of interest

The author declares no conflict of interest. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, and in the decision to publish the results.

Data availability

No data were used to support this study.


  1. Ahanotu E, Alveo-Ceron D, Ravita T, Gaunt E (2006) Staphylococcal enterotoxin B as a biological weapon: recognition management, and surveillance of staphylococcal enterotoxin. Appl Biosaf 11:120–126. CrossRefGoogle Scholar
  2. Argudin MA, Mendoza MC, Rodicio MR (2010) Food poisoning and Staphylococcus aureus enterotoxins. Toxins 2:1751–1773. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Berger T, Eisenkraft A, Bar-Haim E, Kassirer M, Aran AA, Fogel I (2016) Toxins as biological weapons for terror-characteristics, challenges and medical countermeasures: a mini-review. Disaster Mil Med 2:7. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bunroddith K, Viseshakul N, Chansiri K, Lieberzeit P (2018) QCM-based rapid detection of PCR amplification products of Ehrlichia canis. Anal Chim Acta 25:106–111. CrossRefGoogle Scholar
  5. Caughlan MP, Alcock SJ (1991) Chemical sensors for in vivo monitoring: packaging, biocompatibility, stability. Biosens Bioelectron 6:87–91CrossRefGoogle Scholar
  6. de Almeida CC, Pizauro LJL, Soltes GA, Slavic D, de Avila FA, Pizauro JM, MacInnes JI (2018) Some coagulase negative Staphylococcus spp. isolated from buffalo can be misidentified as Staphylococcus aureus by phenotypic and Sa442 PCR methods. BMC Res Notes 11:346. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Dong ZM, Zhao GC (2015) Label-free detection of pathogenic bacteria via immobilized antimicrobial peptides. Talanta 137:55–61. CrossRefPubMedGoogle Scholar
  8. Esposito S et al (2018) Staphylococcus aureus colonization and risk of surgical site infection in children undergoing clean elective surgery: a cohort study. Medicine 97:0000000000011097. CrossRefGoogle Scholar
  9. Fries BC, Varshney AK (2013) Bacterial toxins-staphylococcal enterotoxin B. Microbiol Spectr 1:0002–2012. CrossRefGoogle Scholar
  10. Gubanova O, Andrianova M, Saveliev M, Komarova N, Kuznetsov E, Kuznetsov A (2017) Fabrication and package of ISFET biosensor for micro volume analysis with the use of direct ink writing approach. Mater Sci Semicond Process 60:71–78. CrossRefGoogle Scholar
  11. Guntupalli R et al (2012) Detection and identification of methicillin resistant and sensitive strains of Staphylococcus aureus using tandem measurements. J Microbiol Methods 90:182–191. CrossRefPubMedGoogle Scholar
  12. Guntupalli R, Sorokulova I, Olsen E, Globa L, Pustovyy O, Vodyanoy V (2013) Biosensor for detection of antibiotic resistant Staphylococcus bacteria. J Vis Exp 8:50474. CrossRefGoogle Scholar
  13. Ivbule M, Miklasevics E, Cupane L, Berzina L, Balins A, Valdovska A (2017) Presence of methicillin-resistant Staphylococcus aureus in slaughterhouse environment, pigs, carcasses, and workers. J Vet Res 61:267–277. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Kataby G, Cojocaru M, Prozorov R, Gedanken A (1999) Coating carboxylic acids on amorphous iron nanoparticles. Langmuir 15:1703–1708. CrossRefGoogle Scholar
  15. Krajewski S et al (2014) Bacterial interactions with proteins and cells relevant to the development of life-threatening endocarditis studied by use of a quartz-crystal microbalance. Anal Bioanal Chem 406:3395–3406. CrossRefPubMedGoogle Scholar
  16. Lai LJ, Yang YW, Lin YK, Huang LL, Hsieh YH (2009) Surface characterization of immunosensor conjugated with gold nanoparticles based on cyclic voltammetry and X-ray photoelectron spectroscopy. Colloid Surf B Biointerfaces 68:130–135. CrossRefPubMedGoogle Scholar
  17. Latif U, Qian J, Can S, Dickert FL (2014) Biomimetic receptors for bioanalyte detection by quartz crystal microbalances—from molecules to cells. Sensors 14:23419–23438. CrossRefPubMedGoogle Scholar
  18. Makhneva E, Farka Z, Skladal P, Zajickova L (2018) Cyclopropylamine plasma polymer surfaces for label-free SPR and QCM immunosensing of salmonella. Sens Actuat B Chem 276:447–455. CrossRefGoogle Scholar
  19. Masdor NA, Altintas Z, Tothill IE (2016) Sensitive detection of Campylobacter jejuni using nanoparticles enhanced QCM sensor. Biosens Bioelectron 78:328–336. CrossRefPubMedGoogle Scholar
  20. Mondal B, Ramlal S, Kingston J (2018) Colorimetric DNAzyme biosensor for convenience detection of enterotoxin B harboring Staphylococcus aureus from food samples. J Agric Food Chem 66:1516–1522. CrossRefPubMedGoogle Scholar
  21. Nagai H, Tomioka K, Okumura S (2018) Optimal conditions for the asymmetric polymerase chain reaction for detecting food pathogenic bacteria using a personal SPR sensor. Appl Biochem Biotechnol 26:018–2819. CrossRefGoogle Scholar
  22. Nouri A, Ahari H, Shahbazzadeh D (2018) Designing a direct ELISA kit for the detection of Staphylococcus aureus enterotoxin A in raw milk samples. Int J Biol Macromol 107:1732–1737. CrossRefPubMedGoogle Scholar
  23. Olsson AL, Sharma PK, Mei HC, Busscher HJ (2012) Adhesive bond stiffness of Staphylococcus aureus with and without proteins that bind to an adsorbed fibronectin film. Appl Environ Microbiol 78:99–102. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Pinchuk IV, Beswick EJ, Reyes VE (2010) Staphylococcal enterotoxins. Toxins 2:2177–2197. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Pirich CL, de Freitas RA, Torresi RM, Picheth GF, Sierakowski MR (2017) Piezoelectric immunochip coated with thin films of bacterial cellulose nanocrystals for dengue detection. Biosens Bioelectron 92:47–53. CrossRefPubMedGoogle Scholar
  26. Pizauro LJL et al (2017) Species level identification of coagulase negative Staphylococcus spp from buffalo using matrix-assisted laser desorption ionization-time of flight mass spectrometry and cydB real-time quantitative PCR. Vet Microbiol 204:8–14. CrossRefPubMedGoogle Scholar
  27. Pohanka M (2017a) The piezoelectric biosensors: principles and applications, a review. Int J Electrochem Sci 12:496–506. CrossRefGoogle Scholar
  28. Pohanka M (2017b) The piezoelectric biosensors: principles and applications, a review. Intern J Electrochem Sci 12:496–506. CrossRefGoogle Scholar
  29. Pohanka M (2018a) Overview of piezoelectric biosensors, immunosensors and DNA sensors and their applications. Materials 11:448. CrossRefPubMedCentralGoogle Scholar
  30. Pohanka M (2018b) Piezoelectric biosensor for the determination of tumor necrosis factor alpha. Talanta 178:970–973. CrossRefPubMedGoogle Scholar
  31. Pohanka M, Vlcek V (2018) Assay of glomalin using a quartz crystal microbalance biosensor. Electroanalysis 30:453–458. CrossRefGoogle Scholar
  32. Qudratullah Muhammad G, Saqib M, Bilal MQ (2017) Isolation, characterization, virulence and immunogenicity testing of field isolates of Pasteurella multocida, Staphylococcus aureus, and Streptococcus agalactiae in laboratory settings. Acta Trop 172:70–74. CrossRefPubMedGoogle Scholar
  33. Salam F, Uludag Y, Tothill IE (2013) Real-time and sensitive detection of salmonella Typhimurium using an automated quartz crystal microbalance (QCM) instrument with nanoparticles amplification. Talanta 115:761–767. CrossRefPubMedGoogle Scholar
  34. Shahbazi R, Salouti M, Amini B, Jalilvand A, Naderlou E, Amini A, Shams A (2018) Highly selective and sensitive detection of Staphylococcus aureus with gold nanoparticle-based core-shell nano biosensor. Mol Cell Probes 25:30159. CrossRefGoogle Scholar
  35. Smolensky ED, Park HY, Berquo TS, Pierre VC (2011) Surface functionalization of magnetic iron oxide nanoparticles for MRI applications—effect of anchoring group and ligand exchange protocol. Contrast Media Mol Imaging 6:189–199. CrossRefPubMedGoogle Scholar
  36. Szalontai H, Adanyi N, Kiss A (2014) Comparative determination of two probiotics by QCM and OWLS-based immunosensors. N Biotechnol 31:395–401. CrossRefPubMedGoogle Scholar
  37. Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG Jr (2015) Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 28:603–661. CrossRefPubMedPubMedCentralGoogle Scholar
  38. van den Brand M et al (2018) Evaluation of a real-time PCR assay for detection and quantification of bacterial DNA directly in blood of preterm neonates with suspected late-onset sepsis. Crit Care 22:018–2010. CrossRefGoogle Scholar
  39. Vitko NP, Grosser MR, Khatri D, Lance TR, Richardson AR (2016) Expanded glucose import capability affords Staphylococcus aureus optimized glycolytic flux during infection. MBio 7:00296. CrossRefGoogle Scholar
  40. Wallace SA, Wallace DA, Wood BE (1999) Tests of the sensitivity and mass range of a 50-MHz quartz crystal microbalance (QCM). In: SPIE proceedings 3784:SPIE 3784
  41. Yu S, Chow GM (2004) Carboxyl group (− CO2H) functionalized ferrimagnetic iron oxide nanoparticles for potential bio-applications. J Mater Chem 14:2781–2786CrossRefGoogle Scholar
  42. Zhao Y, Zhu A, Tang J, Tang C, Chen J (2017) Identification and measurement of staphylococcal enterotoxin M from Staphylococcus aureus isolate associated with staphylococcal food poisoning. Lett Appl Microbiol 65:27–34. CrossRefPubMedGoogle Scholar
  43. Zhao X, Liu Z, Meng R, Shi C, Chen X, Bu X, Guo N (2018) Phenotype and RNA-seq-Based transcriptome profiling of Staphylococcus aureus biofilms in response to tea tree oil. Microb Pathog 123:304–313. CrossRefPubMedGoogle Scholar
  44. Zhu L, Zhang Y, He P, Wang Q (2018) A multiplex PCR amplification strategy coupled with microchip electrophoresis for simultaneous and sensitive detection of three foodborne bacteria. J Chromatogr B Analyt Technol Biomed Life Sci 1:1093–1094. CrossRefGoogle Scholar

Copyright information

© Institute of Chemistry, Slovak Academy of Sciences 2019

Authors and Affiliations

  1. 1.Faculty of Military Health SciencesUniversity of DefenceHradec KraloveCzech Republic

Personalised recommendations