DNA PROBE TEST TO DETECT ORAL CANDIDA IN AIDS PATIENTS

THIS RESEARCH WILL PROVIDE THE FOUNDATION FOR A RAPID, INEXPENSIVE, NON-CULTURE-BASED TEST FOR THE QUANTITATIVE DETECTION OF ORAL CANDIDA ALBICANS IN HIV-POSITIVE PATIENTS. THE PRIMARY EXPERIMENTAL OBJECTIVE IS TO CREATE A SET OF SHORT SYNTHETIC OLIGODEOXYNUCLEOTIDE PROBES THAT WILL HYBRIDIZE SPECIFICALLY WITH THE 16S RRNA OF C. ALBICANS. SPECIFICITY WILL BE VERIFIED BY HYBRIDIZATION OF EACH (32)P-LABELED PROBE TO SLOT BLOT FILTERS CONTAINING NUCLEIC ACID SAMPLES FROM OVER 50 MICROORGANISMS THAT ARE FOUND FREQUENTLY IN THE OROPHARYNX. FURTHERMORE, THE (32)-LABELEDPROBES WILL BE USED TO MEASURE LEVELS OF C. ALBICANS IN SAMPLES FROM IMMUNOCOMPROMISED PATIENTS. PROBE RESULTS WILL BE COMPARED WITH CONVENTIONAL DIAGNOSTIC EVALUATIONS OF CANDIDIASIS IN THESE SAME PATIENTS. DURING PHASE II OF THIS PROJECT, THESE PROBES WILL BE USED TO DEVELOP A RAPID COLORIMETRIC ASSAY FOR DETECTING AND MONITORING C. ALBICANS IN PERSONS AT HIGH RISK FOR DEVELOPING ACQUIRED IMMUNODEFICIENCY SYNDROME (AIDS). THE FINAL PRODUCT WILL BE A QUANTITATIVE TEST FOR THE CLINICAL LABORATORY THAT CAN BE PERFORMED WITHIN 3 HOURS OR LESS AT ROOM TEMPERATURE AND THAT CAN BE USED AS AN ALTERNATIVE FOR THE MUCH LENGTHIER PROCEDURE OF CULTURING FOR C. ALBICANS.