Bridge-DNA synthesis triggered by an allosteric aptamer for the colorimetric detection of pathogenic bacteria

Abstract

Rapid and sensitive quantification of pathogenic bacteria is highly desired for environmental health supervision and food safety control. Yet, the amplification and detection of bacteria with a concentration lower than 10² cfu mL⁻¹ remains a great challenge. Here, we combined an allosteric aptamer (AAP) with a gold nanoparticle (AuNP) for assembling a bridge-DNA synthesis system (named as AuNP-BDS) to amplify the bacterial signals. The AAP and its paired primer (PP) were covalently linked to two different AuNPs, respectively: one named as AAP–AuNP and the other PP–AuNP. Upon recognition of the antigen from the pathogenic bacteria, AAP alters its conformation to initiate DNA synthesis on the AuNP surface. The DNA products from AAP–AuNP and PP–AuNP form bridges to each other through base pairing, resulting in the aggregation and colorimetric response of the AuNPs. By using E. coli O157:H7 as an example, the AuNP-BDS could quantify pathogenic bacteria in water with a concentration as low as 10 cfu mL⁻¹ within 60 min and without any enrichment. The colorimetric response values of AuNP-BDS were found to be linearly related to the bacterial concentrations in the range of 10 to 10³ cfu mL⁻¹. Good practicability of the AuNP-BDS in quantifying E. coli O157:H7 from tap water, juices, and milks was demonstrated. The AuNP-BDS could be exploited to facilitate the rapid and sensitive quantification of pathogenic bacteria for food safety control.