Design of a core-shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples

Ozalp V. C., BAYRAMOĞLU G., ARICA M. Y., ÖKTEM H. A.

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, cilt.97, sa.21, ss.9541-9551, 2013 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 97 Sayı: 21
  • Basım Tarihi: 2013
  • Doi Numarası: 10.1007/s00253-013-5231-0
  • Dergi Adı: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.9541-9551
  • Anahtar Kelimeler: Immuno-magnetic separation, Multiplex PCR, Food-borne diseases, Salmonella, Escherichia coli, ESCHERICHIA-COLI O157-H7, SALMONELLA-TYPHIMURIUM, LISTERIA-MONOCYTOGENES, IMMUNOMAGNETIC SEPARATION, BEADS, IMMOBILIZATION, ADSORPTION, LIPASE, CHAIN
  • Orta Doğu Teknik Üniversitesi Adresli: Evet

Özet

We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core-shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) of target bacterial cells from others, antibodies for Escherichia coli and Salmonella enterica serovar Typhimurium cells were immobilized on the magnetic beads via glutaraldehyde coupling reaction. Our IMS system successfully separated Salmonella cells when the concentrations of target (i.e., Salmonella) and interfering (i.e., E. coli) cells were at the same level. Polymerase chain reaction (PCR) assays amplifying the rfb/rfbE region of the E. coli genome and a 647-bp fragment of the invA region of Salmonella were performed as the specific selection to accurately confirm the presence of E. coli and Salmonella, respectively. IMS and multiplex PCR methods can be used for specific and quantitative detection of pathogens from food samples. Thus, this study developed a reliable and direct system for rapid detection of Salmonella and E. coli in food samples. In addition, IMS method could be easily adapted to detect other pathogens by selecting the pertinent antibody.