Simultaneous electrochemical determination of nuc and mecA genes for identification of methicillin-resistant Staphylococcus aureus using N-doped porous carbon and DNA-modified MOF

Mikrochim Acta. 2021 Jan 12;188(2):39. doi: 10.1007/s00604-020-04698-6.

Abstract

The detection of Staphylococcus aureus specific gene in combination with the mecA gene is vitally important for accurate identification of methicillin-resistant Staphylococcus aureus (MRSA). A homogeneous electrochemical DNA sensor was fabricated for simultaneous detection of mecA and nuc gene in MRSA. Metal-organic framework (type UiO-66-NH2) was applied as nanocarrier. Two electroactive dyes, methylene blue (MB) and epirubicin (EP), were encapsulated in UiO-66-NH2, respectively, and were locked by the hybrid double-stranded DNA. Based on the target-response electroactive dye release strategy, once target DNA exists, it completely hybridizes with displacement DNA (DEP and DMB). So DEP and DMB is displaced from the MOF surface, causing the release of electroactive dyes. Co-Zn bimetallic zeolitic imidazolate framework-derived N-doped porous carbon serves for electrode modification to improve electrocatalytic performance and sensitivity. The differential pulse voltammetry peak currents of MB and EP were accurately detected at - 0.14 V and - 0.53 V versus the Ag/AgCl reference electrode, respectively. Under the optimal conditions, the detection limits of mecA gene and nuc gene were 3.7 fM and 1.6 fM, respectively. Combining the effective application of MOFs and the homogeneous detection strategy, the sensor exhibited satisfactory performance for MRSA identification in real samples. The recovery was 92.6-103%, and the relative standard deviation was less than 5%. Besides, MRSA and SA can also be distinguished. This sensor has great potential in practical applications.

Keywords: Co-Zn bimetallic ZIF; Differential pulse voltammetry; Electrochemical DNA sensor; Foodborne bacteria; Modified electrode; UiO-66.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods
  • Carbon / chemistry*
  • Coloring Agents / chemistry
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Drinking Water / analysis
  • Drinking Water / microbiology
  • Electrochemical Techniques / instrumentation
  • Electrochemical Techniques / methods*
  • Electrodes
  • Epirubicin / chemistry
  • Immobilized Nucleic Acids / chemistry*
  • Immobilized Nucleic Acids / genetics
  • Limit of Detection
  • Metal-Organic Frameworks / chemistry*
  • Methicillin-Resistant Staphylococcus aureus / chemistry*
  • Methylene Blue / chemistry
  • Micrococcal Nuclease / genetics
  • Milk / microbiology
  • Nucleic Acid Hybridization
  • Organometallic Compounds / chemistry
  • Penicillin-Binding Proteins / genetics
  • Phthalic Acids / chemistry
  • Reproducibility of Results

Substances

  • Bacterial Proteins
  • Coloring Agents
  • DNA, Bacterial
  • Drinking Water
  • Immobilized Nucleic Acids
  • Metal-Organic Frameworks
  • Organometallic Compounds
  • Penicillin-Binding Proteins
  • Phthalic Acids
  • UiO-66
  • mecA protein, Staphylococcus aureus
  • nuc protein, staphylococcus
  • Epirubicin
  • Carbon
  • Micrococcal Nuclease
  • Methylene Blue