Electrochemical Detection for Isothermal Loop-Mediated Amplification of Pneumolysin Gene of Streptococcus pneumoniae Based on the Oxidation of Phenol Red Indicator

Anal Chem. 2022 Sep 27;94(38):13061-13067. doi: 10.1021/acs.analchem.2c02127. Epub 2022 Sep 15.

Abstract

A highly sensitive electrochemical methodology for end-point detection of loop-mediated isothermal nucleic acid amplification reactions was developed. It is based on the oxidation process of phenol red (PR), commonly used as a visual indicator. The dependence of its redox process on pH, which changes during amplification, allows performing quantitative measurements. Thus, the change in the oxidation potential of PR during the amplification is used, for the first time, as the analytical signal that correlates with the number of initial DNA copies. As a proof-of-concept, the amplification of the pneumolysin gene from Streptococcus pneumoniae, one of the main pathogens causing community-acquired pneumonia, is performed. Combination of isothermal amplification with electrochemical detection, performed on small-size flexible electrodes, allows easy decentralization. Adaptation to the detection of other pathogens causing infectious diseases would be very useful in the prevention of future epidemics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins
  • DNA
  • Electrochemical Techniques
  • Nucleic Acid Amplification Techniques / methods
  • Oxidation-Reduction
  • Phenolsulfonphthalein*
  • Streptococcus pneumoniae* / genetics
  • Streptolysins

Substances

  • Bacterial Proteins
  • Streptolysins
  • plY protein, Streptococcus pneumoniae
  • DNA
  • Phenolsulfonphthalein