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Characterization of oral swab samples for diagnosis of pulmonary tuberculosis.

ABSTRACT: Oral swab analysis (OSA) has been shown to detect Mycobacterium 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.

SUBMITTER: Wood RC

PROVIDER: S-EPMC8128230 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Characterization of oral swab samples for diagnosis of pulmonary tuberculosis.

Wood Rachel C RC Andama Alfred A Hermansky Gleda G Burkot Stephen S Asege Lucy L Job Mukwatamundu M Katumba David D Nakaye Martha M Mwebe Sandra Z SZ Mulondo Jerry J Bachman Christine M CM Nichols Kevin P KP Le Ny Anne-Laure M AM Ortega Corrie C Olson Rita N RN Weigel Kris M KM Olson Alaina M AM Madan Damian D Bell David D Cattamanchi Adithya A Worodria William W Semitala Fred C FC Somoskovi Akos A Cangelosi Gerard A GA Minch Kyle J KJ

PloS one 20210517 5

Oral swab analysis (OSA) has been shown to detect Mycobacterium 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 r ...[more]

PMID: 33999938

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Project description:ObjectivesTuberculosis (TB) remains a significant concern in terms of public health, necessitating the timely and accurate diagnosis to impede its advancement. The utilization of oral swab analysis (OSA) presents a promising approach for diagnosing pulmonary TB by identifying Mycobacterium tuberculosis (MTB) within oral epithelial cells. Due to disparities in the diagnostic performance of OSA reported in the original studies, we conducted a meticulous meta-analysis to comprehensively assess the diagnostic efficacy of OSA in pulmonary TB.MethodsWe conducted a comprehensive investigation across multiple databases, namely PubMed, Cochrane Library, Embase, Web of Science, ClinicalTrials.gov, Chinese BioMedical Literature Database (CBM), China National Knowledge Infrastructure Database (CNKI), and Wanfang China Science and Technology Journal Database to identify relevant studies. Out search query utilized the following keywords: oral swab, buccal swab, tongue swab, tuberculosis, and TB. Subsequently, we employed STATA 16.0 to compute the combined sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio for both the overall and subgroup analyses.ResultsOur findings indicated that OSA has a combined sensitivity of 0.67 and specificity of 0.95 in individuals with pulmonary TB. Subgroup analysis further revealed that among adult individuals with pulmonary TB, the sensitivity and specificity of OSA were 0.73 and 0.93, respectively. In HIV-negative individuals with pulmonary TB, the sensitivity and specificity were 0.68 and 0.98, respectively. The performance of OSA in detecting pulmonary TB correlated with the bacteria load in sputum. Additionally, the sensitivity for diagnosing pulmonary TB using tongue specimens was higher (0.75, 95% CI: 0.65-0.83) compared to cheek specimens (0.52, 95% CI: 0.34-0.70), while both types of specimens demonstrated high specificity.ConclusionsTo conclude, oral swabs serve as a promising alternative for diagnosing pulmonary TB, especially in adult patients. In addition, tongue swabs yield better sensitivity than cheek swabs to identify pulmonary TB patients.Systematic review registrationidentifier: CRD42023421357.

Project description:Tongue swab (TS) sampling combined with quantitative PCR (qPCR) to detect Mycobacterium tuberculosis (MTB) DNA is a promising alternative to sputum testing for tuberculosis (TB) diagnosis. In prior studies, the sensitivity of tongue swabbing has usually been lower than sputum. In this study, we evaluated two strategies to improve sensitivity. In one, centrifugation was used to concentrate tongue dorsum bacteria from 2-mL suspensions eluted from high-capacity foam swab samples. The pellets were resuspended as 500-µL suspensions, and then mechanically lysed prior to dual-target qPCR to detect MTB insertion elements IS6110 and IS1081. Fractionation experiments demonstrated that most of the MTB DNA signal in clinical swab samples (99.22% ± 1.46%) was present in the sedimentable fraction. When applied to archived foam swabs collected from 124 South Africans with presumptive TB, this strategy exhibited 83% sensitivity (71/86) and 100% specificity (38/38) relative to sputum microbiological reference standard (MRS; sputum culture and/or Xpert Ultra). The second strategy used sequence-specific magnetic capture (SSMaC) to concentrate DNA released from MTB cells. This protocol was evaluated on archived Copan FLOQSwabs flocked swab samples collected from 128 South African participants with presumptive TB. Material eluted into 500 µL buffer was mechanically lysed. The suspensions were digested by proteinase K, hybridized to biotinylated dual-target oligonucleotide probes, and then concentrated ~20-fold using magnetic separation. Upon dual-target qPCR testing of concentrates, this strategy exhibited 90% sensitivity (83/92) and 97% specificity (35/36) relative to sputum MRS. These results point the way toward automatable, high-sensitivity methods for detecting MTB DNA in TS.ImportanceImproved testing for tuberculosis (TB) is needed. Using a more accessible sample type than sputum may enable the detection of more cases, but it is critical that alternative samples be tested appropriately. Here, we describe two new, highly accurate methods for testing tongue swabs for TB DNA.

Project description:Tongue swab (TS) sampling combined with qPCR to detect Mycobacterium tuberculosis (MTB) DNA is a promising alternative to sputum testing for tuberculosis (TB) diagnosis. In prior studies, the sensitivity of tongue swabbing has usually been lower than sputum. In this study, we evaluated two strategies to improve sensitivity. In one, centrifugation was used to concentrate tongue dorsum bacteria from 2-mL suspensions eluted from high-capacity foam swab samples. The pellets were resuspended as 500-μL suspensions, and then mechanically lysed prior to dual-target qPCR to detect MTB insertion elements IS6110 and IS1081. Fractionation experiments demonstrated that most of the MTB DNA signal in clinical swab samples (99.22% ± 1.46%) was present in the sedimentable fraction. When applied to archived foam swabs collected from 124 South Africans with presumptive TB, this strategy exhibited 83% sensitivity (71/86) and 100% specificity (38/38) relative to sputum MRS (microbiological reference standard; sputum culture and/or Xpert® Ultra). The second strategy used sequence-specific magnetic capture (SSMaC) to concentrate DNA released from MTB cells. This protocol was evaluated on archived Copan FLOQSwabs® flocked swab samples collected from 128 South African participants with presumptive TB. Material eluted into 500 μL buffer was mechanically lysed. The suspensions were digested by proteinase K, hybridized to biotinylated dual-target oligonucleotide probes, and then concentrated ~20-fold using magnetic separation. Upon dual-target qPCR testing of concentrates, this strategy exhibited 90% sensitivity (83/92) and 97% specificity (35/36) relative to sputum MRS. These results point the way toward automatable, high-sensitivity methods for detecting MTB DNA in TS.ImportanceImproved testing for tuberculosis (TB) is needed. Using a more accessible sample type than sputum may enable the detection of more cases, but it is critical that alternative samples be tested appropriately. Here, we describe two new, highly accurate methods for testing tongue swabs for TB DNA.

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Characterization of oral swab samples for diagnosis of pulmonary tuberculosis.

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Characterization of oral swab samples for diagnosis of pulmonary tuberculosis.

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