1. Characterization of oral swab samples for diagnosis of pulmonary tuberculosis
- Author
-
William Worodria, Adithya Cattamanchi, Akos Somoskovi, David Katumba, Kyle J. Minch, Christine Bachman, Burkot Stephen Thomas Graves, Derek Bell, Anne-Laure M. Le Ny, Rachel C. Wood, Damian Madan, Fred C. Semitala, Kris M. Weigel, Corrie Ortega, Gleda Hermansky, Lucy Asege, Sandra Mwebe, Gerard A. Cangelosi, Rita N. Olson, Alfred Andama, Alaina M. Olson, Kevin Paul Flood Nichols, Jerry Mulondo, Martha Nakaye, Mukwatamundu Job, and Subbian, Selvakumar
- Subjects
Bacterial Diseases ,RNA viruses ,Male ,Physiology ,Pathology and Laboratory Medicine ,Polymerase Chain Reaction ,0302 clinical medicine ,Medical Conditions ,Immunodeficiency Viruses ,Medicine and Health Sciences ,Uganda ,030212 general & internal medicine ,DNA extraction ,Lung ,Multidisciplinary ,GeneXpert MTB/RIF ,biology ,Bacterial ,Pulmonary ,Body Fluids ,Bacterial Pathogens ,Actinobacteria ,RNA, Bacterial ,medicine.anatomical_structure ,Infectious Diseases ,Medical Microbiology ,Viral Pathogens ,Viruses ,Medicine ,Tuberculosis Diagnosis and Management ,HIV/AIDS ,Female ,Sample collection ,medicine.symptom ,Anatomy ,Pathogens ,Infection ,Research Article ,Adult ,Bacilli ,Tuberculosis ,Adolescent ,General Science & Technology ,Science ,030231 tropical medicine ,Microbiology ,DNA, Ribosomal ,Specimen Handling ,Mycobacterium tuberculosis ,Vaccine Related ,03 medical and health sciences ,Young Adult ,Extraction techniques ,Rare Diseases ,Tongue ,Diagnostic Medicine ,Clinical Research ,Retroviruses ,medicine ,Humans ,Dental/Oral and Craniofacial Disease ,Microbial Pathogens ,Tuberculosis, Pulmonary ,Ribosomal ,Mouth ,Bacteria ,business.industry ,Lentivirus ,Sputum ,Organisms ,Biology and Life Sciences ,HIV ,Mycobacteria ,DNA ,biology.organism_classification ,medicine.disease ,Tropical Diseases ,Research and analysis methods ,Mucus ,Genes ,Genes, Bacterial ,RNA, Ribosomal ,RNA ,business ,Digestive System - Abstract
Oral swab analysis (OSA) has been shown to detectMycobacterium tuberculosis(MTB) DNA in patients with pulmonary tuberculosis (TB). In previous analyses, qPCR testing of swab samples collected from tongue dorsa was up to 93% sensitive relative to sputum GeneXpert, when 2 swabs per patient were tested. The present study modified sample collection methods to increase sample biomass and characterized the viability of bacilli present in tongue swabs. A qPCR targeting conserved bacterial ribosomal rRNA gene (rDNA) sequences was used to quantify bacterial biomass in samples. There was no detectable reduction in total bacterial rDNA signal over the course of 10 rapidly repeated tongue samplings, indicating that swabs collect only a small portion of the biomass available for testing. Copan FLOQSwabs collected ~2-fold more biomass than Puritan PurFlock swabs, the best brand used previously (p = 0.006). FLOQSwabs were therefore evaluated in patients with possible TB in Uganda. A FLOQSwab was collected from each patient upon enrollment (Day 1) and, in a subset of sputum GeneXpert Ultra-positive patients, a second swab was collected on the following day (Day 2). Swabs were tested for MTB DNA by manual IS6110-targeted qPCR. Relative to sputum GeneXpert Ultra, single-swab sensitivity was 88% (44/50) on Day 1 and 94.4% (17/18) on Day 2. Specificity was 79.2% (42/53). Among an expanded sample of Ugandan patients, 62% (87/141) had colony-forming bacilli in their tongue dorsum swab samples. These findings will help guide further development of this promising TB screening method.
- Published
- 2021