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Reviews of "High-Sensitivity Detection of Mycobacterium Tuberculosis DNA in Tongue Swab Samples"

Reviewers: M Nielsen (Boston University) | 📗📗📗📗◻️ • L Davis (Yale) | 📘📘📘📘📘

Published onOct 25, 2024
Reviews of "High-Sensitivity Detection of Mycobacterium Tuberculosis DNA in Tongue Swab Samples"
key-enterThis Pub is a Review of
High-sensitivity detection of Mycobacterium tuberculosis DNA in tongue swab samples
High-sensitivity detection of Mycobacterium tuberculosis DNA in tongue swab samples
Description

Abstract 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.Importance Improved 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.

To read the original manuscript, click the link above.

Summary of Reviews: This preprint explores the sensitivity of potential alternative mycobacterium tuberculosis (MTB) diagnostic methods using tongue swabs as an alternative from the current method of testing sputum samples. The reviewers praised the study for demonstrating the potential of oral swabs in TB testing, which is a much easier way to collect samples, allowing for enhanced diagnostic potential of TB. They also praised the preprint for providing detailed methodology that could improve its sensitivity and performance if used in real-world settings. The reviewers had concerns that the study was still preliminary and remains a "proof-of-concept" rather than a clear path forward with feasibility of implementing tongue swabs as the methodologies suggested are not accessible for point of care settings. More studies were recommended for additional comparisons of these methods as well as acquiring samples in real time rather than analyzing archived samples retrospectively.

Reviewer 1 (Marisa N…) | 📗📗📗📗◻️

Reviewer 2 (Lucian (Luke) D…) | 📘📘📘📘📘

RR\ID Strength of Evidence Scale Key

📕 ◻️◻️◻️◻️ = Misleading

📙📙 ◻️◻️◻️ = Not Informative

📒📒📒 ◻️◻️ = Potentially Informative

📗📗📗📗◻️ = Reliable

📘📘📘📘📘 = Strong

To read the reviews, click the links below. 

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