DIAGNOSIS OF TUBERCULOSIS BY LUCIFERASE PHAGE

Project: Research project

Project Details

Description

DESCRIPTION (Adapted from applicant's abstract):Tuberculosis is a
devastating disease of mankind resulting in 3 million deaths per year.
There are 10 million new cases of tuberculosis worldwide, and after a 32-
year decline, the number of new cases is now increasing dramatically in
the United States with a nearly 10 percent increase in the number of AIDS
patients since HIV both reduces immunity against Mycobacteria tuberculosis
infection and activates existing, latent infections. Although there are
effective chemotherapeutic approaches for tuberculosis, rapid diagnosis is
important for effective treatment and current methods take from two to six
weeks. Moreover,there are increasing reports of the apparent appearance
of drug resistantstrains of M. tuberculosis and the determination of drug
susceptibilitiessometimes takes too long to be of use to the patient. The
goal of this project is to develop a rapid, sensitive and specific tool
for the diagnosis of tuberculosis and rapid assessment of drug
sensitivities, using diagnostic reporter bacteriophages.

Diagnostic reporter phages will be constructed by insertion of genes
encoding luciferase enzymes expressed from mycobacterial-specific signals
into mycobacteriophage genomes. When these recombinant phage infect
mycobacteria, the luciferase enzyme is expressed and can be detected with
an extremely sensitive assay for the production of photons in a
luminometer. This assay is very sensitive and should be able to detect
fewer than 100 live M. tuberculosis bacilli in a clinical sample. The
need for cultivation is thus eliminated and diagnosis can be performed in
a few hours rather than a few weeks. Moreover, the response of
antibiotics can be monitored at the same time. Specificity of the assay
will be generated through the construction of several reporter phages that
have different, but well defined host ranges. The phages used will include
the well-characterized L5, TM4, and a number of novel uncharacterized
phages. Phage L5 offers particular advantages as a model for gene
expression following phage infection and for the construction of
alternative cosmid vectors.
StatusFinished
Effective start/end date1/1/908/31/95

Funding

  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases

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