Emerging and re-emerging bacterial diseases threaten the health of the U.S. population and molecular microbiological techniques afford the opportunity for efficient and cost-effective identification of these organisms. Much of the previous work has relied upon unique primers specific to each organism. The investigators propose to amplify bacterial DNA with universal polymerase chain reaction (PCR) primers to highly conserved regions of the 16S ribosomal RNA coding (16S rDNA) sequences, followed by specific probes for variable regions between the universal primer sites. The investigators hypothesize that this approach will be 1) applicable to a broad range of bacteria incuding emerging/re-emerging organisms; 2) more rapid than culture-based methods; and 3) sufficiently sensitive and specific to detect these bacteria in biological samples and water quality samples. To test these hypotheses, the investigators propose the following Specific Aims: 1) to determine whether 16S rDNA universal primers and species specific probes will be applicable to a broad range of organisms, requiring sequencing of this region for organisms in which these data are not yet available; 2) to evaluate the relative speed of the molecular approach compared to culture-based methods; and 3) to determine whether the sensitivity and specificity of this approach will be sufficient to detect and identify a specific organism in samples such as body fluids and drinking water. The investigators speculate that the efficiency and cost-effectiveness of the universal primer amplification for specific bacterial identification will improve diagnosis and surveillance for a broad range of bacteria, including emerging and remerging organisms.
Thesaurus Terms:method development, microorganism classification, polymerase chain reaction Mycobacterium tuberculosis, Rickettsiales, Vibrio choleraeNational Institute of Allergy and Infectious Diseases (NIAID)
