Effects of pretreatments on detection of E. coli O157

H7 by SPR Biosensor

Xiaoguang Zhang, Sachiko Tsuji, Hayato Kitaoka, Mitsuru Tamai, Hiroshi Kobayashi, Ken-ichi Honjoh, Takahisa Miyamoto

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Escherichia coli (E. coli) O157:H7 was detected by using a surface plasmon resonance (SPR) biosensor and two antibodies with different characters. The lower limit of detection of E. coli 0157:H7 samples after pretreatments was determined by SPR. Seven pretreatment methods for preparing E. coli 0157:H7 samples for SPR detection; beads disruption, sonication, and heat shock, osmotic shock, lysozyme, alkali, and boiling treatments were compared for SPR signal with untreated cells as a control. In the case of the antibody raised against intracellular substance, β-D-galactosidase (β-gal), was used for the detection, the lower limit of detection was 4.9x105 CFU/ml for both sonicated and alkali treated samples. The lower limit of detection was 8.3x106 CFU/ml for beads disrupted samples, and 8.2x108 CFU/ml for both lysozyme treated and untreated samples. In contrast, significant SPR signal was not obtained for heat shocked, osmotic shocked and boiled samples even at 8.2xl08 CFU/ml. Sonication pretreatment improved the lower limit of detection for E. coli 0157:H7 by three orders of magnitude compared with that of untreated sample when anti-β-gal antibody was used for detection by SPR biosensor. In the case of antibody raised against lipopolysaccharide (LPS), the cell surface substance, was used, sonicated E. coli 0157:H7 sample was detected by SPR at 1.3xl05 CFU/ml. The lower limit of detection was 1.1x105 CFU/ml for heat shocked, lysozyme treated, alkali treated, boiled and untreated samples, and 7.7x10' CFU/ml for beads disrupted samples, respectively. After the osmotic shock treatment, E. coli O157:H7 was not detected by SPR even at 2.1x108 CFU/ml. These results show that sonication was the most effective pretreatment method for the detection of E. coli O157:H7 by SPR using both antibodies recognizing intracellular β-gal, and cell surface LPS.

Original languageEnglish
Pages (from-to)307-312
Number of pages6
JournalJournal of the Faculty of Agriculture, Kyushu University
Volume58
Issue number2
Publication statusPublished - Sep 1 2013

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surface plasmon resonance
Escherichia coli O157
Surface Plasmon Resonance
biosensors
Biosensing Techniques
pretreatment
detection limit
Limit of Detection
Sonication
Alkalies
Muramidase
sampling
Escherichia coli
antibodies
lysozyme
alkalis
Antibodies
Hot Temperature
Osmotic Pressure
osmotic stress

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Agronomy and Crop Science

Cite this

Effects of pretreatments on detection of E. coli O157 : H7 by SPR Biosensor. / Zhang, Xiaoguang; Tsuji, Sachiko; Kitaoka, Hayato; Tamai, Mitsuru; Kobayashi, Hiroshi; Honjoh, Ken-ichi; Miyamoto, Takahisa.

In: Journal of the Faculty of Agriculture, Kyushu University, Vol. 58, No. 2, 01.09.2013, p. 307-312.

Research output: Contribution to journalArticle

Zhang, Xiaoguang ; Tsuji, Sachiko ; Kitaoka, Hayato ; Tamai, Mitsuru ; Kobayashi, Hiroshi ; Honjoh, Ken-ichi ; Miyamoto, Takahisa. / Effects of pretreatments on detection of E. coli O157 : H7 by SPR Biosensor. In: Journal of the Faculty of Agriculture, Kyushu University. 2013 ; Vol. 58, No. 2. pp. 307-312.
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