Food and Fermentation Industries >

2022 , Vol. 48 >Issue 2: 274 - 279

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027618

Colorimetric detection of Staphylococcus aureus using aptamers and hybridization chain reaction amplification

  • LU Chunxia ,
  • WANG Changxuan ,
  • HUANG Qiao ,
  • CHEN Xia ,
  • LI Hongmin ,
  • FU Jiali ,
  • ZHANG Yunhong ,
  • WANG Shuanghui
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  • 1(School of Advanced Agriculture and Bioengineering, Yangtze Normal University, Chongqing 408100, China)
    2(Suzhou Saifeford Testing Technology Co.Ltd., Suzhou 215000, China)
    3(Analysis and Testing Center, Xinjiang Academy of Agriculture and Reclamation Science, Shihezi 832000, China)

Received date: 2021-04-05

  Revised date: 2021-05-04

  Online published: 2022-02-28

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Abstract

A ultrasensitive colorimetric assay was developed for the detection of Staphylococcus aureus based on hybridization chain reaction with enzyme-amplification using aptamer as the recognition probe. The results showed that the optimal conditions for this method were as follows: aptamer 60 nmol/L, detection probe 80 nmol/L, hairpin DNA 80 nmol/L, bovine serum albumin concentration 10 g/L, blocking time 4 h, dilution volume ratio of streptavidin-horseradish peroxidase (SA-HRP) 1:40 000, incubation time between S. aureus and aptamers 30 min, hybridization time of hairpin DNA 60 min. Under the optimal conditions, the calibration curves for S. aureus showed good linearities in the range of 10-105 CFU/mL with correlation coefficients (R) of 0.992. The limit of detection was 10 CFU/mL, and there was no cross-reaction with other food-borne pathogens. Subsequently, the proposed method was applied to measure S. aureus in real samples, and was validated using official standard method. There was no significant difference between the detection results of the two methods (P > 0.05). The recoveries of S. aureus in food samples were in the range of 91.8%-98.2%, and the relative standard deviation was less than 6%. This developed method has the advantages of simple pretreatment, high sensitivity, low detection cost and good accuracy, and is suitable for rapid determination of S. aureus in batch samples.

Key words： Staphylococcus aureus; aptamer; hybridization chain reaction; colorimetric assay

Cite this article

LU Chunxia , WANG Changxuan , HUANG Qiao , CHEN Xia , LI Hongmin , FU Jiali , ZHANG Yunhong , WANG Shuanghui . Colorimetric detection of Staphylococcus aureus using aptamers and hybridization chain reaction amplification[J]. Food and Fermentation Industries, 2022 , 48(2) : 274 -279 . DOI: 10.13995/j.cnki.11-1802/ts.027618

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