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David Geffen School of Medicine at University of California Los Angeles

2 ARTICLES PUBLISHED IN JoVE

Bioengineering

Bacterial Detection & Identification Using Electrochemical Sensors

Colin Halford^1,2, Vincent Gau³, Bernard M. Churchill², David A. Haake^2,4,5

¹Research Service, Veterans Affairs Greater Los Angeles Healthcare System, ²Department of Urology, The David Geffen School of Medicine, University of California, Los Angeles , ³GeneFluidics, ⁴Division of Infectious Diseases, Veterans Affairs Greater Los Angeles Healthcare System, ⁵Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles

We describe an electrochemical sensor assay method for rapid bacterial detection and identification. The assay involves a sensor array functionalized with DNA oligonucleotide capture probes for ribosomal RNA (rRNA) species-specific sequences. Sandwich hybridization of target rRNA with the capture probe and a horseradish peroxidase-linked DNA oligonucleotide detector probe produces a measurable amperometric current.

Immunology and Infection

High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection

Kristel Lourdault^1,2, James Matsunaga^1,2, Karen V. Evangelista^1,2, David A. Haake^1,2,3,4

¹Veterans Affairs Greater Los Angeles Healthcare System, ²Departments of Medicine, David Geffen School of Medicine at University of California Los Angeles, ³Departments of Urology, David Geffen School of Medicine at University of California Los Angeles, ⁴Departments of Microbiology, Immunology, and Molecular Genetics, University of California Los Angeles

We describe here a technique that combines transposon mutagenesis with high-throughput sequencing to identify and quantify transposon leptospiral mutants in tissues after a challenge of hamsters. This protocol can be used to screen mutants for survival and dissemination in animals and can also be applied to in vitro studies.