Veterans Affairs Greater Los Angeles Healthcare System

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Immuno-fluorescence Assay of Leptospiral Surface-exposed Proteins

¹Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, ²Research service, 151, Veterans Affairs Greater Los Angeles Healthcare System, ³Departments of Medicine, Urology at David Geffen School of Medicine and Department of Microbiology, Immunology and Molecular Gentics, University of California Los Angeles (UCLA), ⁴Division of Infectious Diseases, 111F, Veterans Affairs Greater Los Angeles Health Care System

An efficient method to assess surface-exposure of leptospiral proteins is described. The method is specifically designed to avoid disruption of the fragile outer membrane of leptospiral cells. This technique requires employment of several negative controls to assess the integrity of the outer membrane and specificity of antibody reaction.

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.