Lateral Flow Hybridization Assay to Determine Transcripts of TEM-Type Beta-Lactamase Genes in Bacteria Resistant to Antibiotics

Abstract

A technique for lateral flow hybridization analysis of specific bacterial beta-lactamase mRNAs, including the preparation of a biotinylated target DNA and its analysis on a test strip with immobilized oligonucleotide probes, is developed. When the target DNA and probe structures are complementary, DNA duplexes are formed in the test zone of the strip, which interact with the conjugate of streptavidin and gold nanoparticles. The technique is used to determine the transcripts of the genes encoding TEM-type beta-lactamases isolated from the culture of E. coli producing the corresponding recombinant beta-lactamase. It is shown that single-stranded 330 bp target DNA is determined most effectively in lateral flow analysis.