Antimicrobial resistance and bacterial identification utilizing a microelectronic chip array

Species-specific bacterial identification of clinical specimens is often li mited to a few species due to the difficulty of performing multiplex reacti ons. In addition, discrimination of amplicons is time-consuming and laborio us, consisting of gel electrophoresis, probe hybridization, or sequencing t echnology. In order to simplify the process of bacterial identification, we combined anchored in situ amplification on a microelectronic chip array wi th discrimination and detection on the same platform. Here, we describe the simultaneous amplification and discrimination of six gene sequences which are representative of different bacterial identification assays: Escherichi a coli gyrA, Salmonella gyrA, Campylobacter gyrA, E. coli parC, Staphylococ cus mecA, and Chlamydia cryptic plasmid. The assay can detect both plasmid and transposon genes and can also discriminate strains carrying antibiotic resistance single-nucleotide polymorphism mutations. Finally, the assay is similarly capable of discriminating between bacterial species through repor ter-specific discrimination and allele-specific amplification. Anchored str and displacement amplification allows multiplex amplification and complex g enotype discrimination on the same platform. This assay simplifies the bact erial identification process greatly, allowing molecular biology techniques to be performed with minimal processing of samples and practical experienc e.