Fast fluorometric enumeration of E. coli using passive chip


KASAP E. N., DOĞAN Ü., ÇOĞUN F., YILDIRIM E., BOYACI İ. H., ÇETİN D., ...Daha Fazla

JOURNAL OF MICROBIOLOGICAL METHODS, cilt.164, 2019 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 164
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.mimet.2019.105680
  • Dergi Adı: JOURNAL OF MICROBIOLOGICAL METHODS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Anahtar Kelimeler: E. coil, Microfluidic chip, Immunornagnetic separation, Fluorescence, Sandwich immunoassay, ESCHERICHIA-COLI, QUANTUM DOTS, RAPID DETECTION, IMMUNOMAGNETIC SEPARATION, IMMUNOSENSOR, FABRICATION, BIOSENSOR, BACTERIA, O157-H7, PROBE
  • Orta Doğu Teknik Üniversitesi Adresli: Evet

Özet

In this report, a passive microfluidic chip design was developed for fast and sensitive fluorometric determination of Escherichia coli (E. coli) based on sandwich immunoassay. Initially, magnetic nanoparticles (MNPs) and chitosan modified mercaptopropionic acid capped cadmium telluride (CdTe) quantum dots (QDs) were functionalized with E.coli specific antibody to form a sandwich immunoassay with the E. coli. The magnetic separation and preconcentration of the E.coli from the sample solution was performed in the vial. Conjugation of QDs to the magnetically captured E. coli and washing were performed using a passive type of microchip. The microfluidic chip consists of four microchambers connected to each other by microchannels which act as capillary valves. Signal measurement was performed at the last chamber by using a hand-held spectrofluorometer equipped with a fiber optic reflection probe. The selectivity of the method was tested with Enterobacter aerogenes (E. aerogenes) and Salmonella enteritidis (S. enteritidis), it was observed that these bacteria have no interference effect on E.coli determination. The calibration curve was found to be linear in the range of 10(1)-10(5) cfu/mL with a correlation coefficient higher than 0.99. The limit of detection was calculated as 5 cfu/mL. The method was successfully applied to spiked tap and lake water samples. The results suggest that the developed method is applicable for on-site E. coli detection and offers several advantages such as large dynamic range, high sensitivity, high selectivity and short analysis time.