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| Grant Number: | 1R41AI052523-01 |
| PI Name: | ALLAND, DAVID E. |
| PI Email: | dalland404@aol.com |
| PI Title: | ASSOCIATE PROFESSOR OF MEDICINE |
| Project Title: | Sample Processing Cartridges for Rapid PCR TB Detection |
Abstract: DESCRIPTION (provided by applicant): Conventional methods to identify and test the antibiotic susceptibility of Mycobacterium tuberculosis are both slow and insensitive. Current nucleic acid amplification-based techniques present a partial solution by offering a more rapid result, but these methods require a relatively high degree of technical sophistication. Furthermore, assay sensitivity is often limited because there are no reliable methods either to concentrate the bacilli within a sputum sample into a small volume for polymerase chain reaction (PCR) amplification, or to extract M. tuberculosis DNA from sputum samples that is free of PCR-reaction inhibitors. We have developed an extremely sensitive and specific single-tube PCR assay that identifies M. tuberculosis and detects resistance to rifampin directly from clinical sputum samples. In this phase I STIR, we will develop a low-cost cartridge-based PCR system that will combine dramatically increased sensitivity with completely "hands-free" operation and analysis. The specific aims are 1) to develop a low-cost cartridge system that will concentrate all of the M. tuberculosis bacilli within an entire sputum sample into a small volume that can be used in a single PCR reaction, then wash the concentrated cells free of PCR inhibitors and completely lyse the cells; 2) to develop a whole-cell mycobacterial reporter that will serve as a quality control for all assay steps from sputum processing through PCR and analysis; and 3) to test the system using clinical sputum specimens. This project will result in a rapid system that is sensitive and specific enough to be performed on patients with smear-negative tuberculosis, and that is simple and inexpensive enough be performed almost anywhere with minimal training.
Thesaurus Terms:
Mycobacterium tuberculosis, biomedical equipment development, communicable disease diagnosis, diagnosis design /evaluation, rapid diagnosis, sample collection
drug resistance, method development, rifamycin, sputum
clinical research, human tissue, polymerase chain reaction
| Institution: | CEPHEID |
| 1190 BORREGAS AVE | |
| SUNNYVALE, CA 94089 | |
| Fiscal Year: | 2002 |
| Department: | |
| Project Start: | 15-AUG-2002 |
| Project End: | 14-AUG-2003 |
| ICD: | NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES |
| IRG: | ZRG1 |