Rapid and label-free detection of Brucella melitensis in milk and milk products using an aptasensor

BAYRAMOĞLU G., Ozalp V. C., Oztekin M., Arica M. Y.

TALANTA, cilt.200, ss.263-271, 2019 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 200
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.talanta.2019.03.048
  • Dergi Adı: TALANTA
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.263-271
  • Anahtar Kelimeler: Magnetic nanoparticle, QCM, Aptamer, Aptasensor, Brucella melitensis, Milk and milk products, QUARTZ-CRYSTAL MICROBALANCE, BIOLOGICAL WARFARE, PATHOGEN DETECTION, BIOSENSOR, IDENTIFICATION, SALMONELLA, DESIGN, LIGAND, QCM, IMMOBILIZATION
  • Gazi Üniversitesi Adresli: Evet

Özet

In this work, a novel quartz crystal microbalance (QCM) aptasensor is designed for the diagnosis of Brucella melitensis bacteria, which affects the Mediterranean fever (brucellosis) from the zoonotic diseases that are very common in the Middle East Countries. The method is based on the selection of B. melitensis bacterium from solutions using B. melitensis specific binding aptamer (Apt) attached magnetic nanoparticles. The surface of the magnetic nanoparticles (i.e.,Fe3O4) was modified by 3-aminopropyltriethoxysilane (APTES) and then grafted with a hydrophilic macromonomer poly(ethyleneglycol)-methacrylate (PEG-MA) as a first block polymer and glycidylinethacrylate (GMA) as a second block functional polymer via atom transfer radical polymerization (ATRP) method [Fe3O4 @SiO2 @p(PEG-MA-GMA)], then, the specific binding aptamer was immobilized. The aptamer immobilized magnetic nanoparticles were used for the pre-concentration of the target bacterium, and the same aptamer sequence was also immobilized on the QCM chip and used for the quantitative detection of B. melitensis using QCM aptasensor. The detection limits of the QCM aptasensor were in the range 1.0(2)-1.0(7) CFU mL(-1), with recoveries up to 79%. The synthesized [Fe3O4 @SiO2 @P(PEGMA-GMA)] nanoparticles showed a good permanence and high isolation recoveries for the pull down of the target bacterium from food samples, after recycling eight times. The method was successfully applied to target bacterium determinations in milk and milk product samples.