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. 2023 Jan 12;12:1064148. doi: 10.3389/fcimb.2022.1064148

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Copyright © 2023 Peters, McIvor, Papadopoulos, Masangana, Gordhan, Waja, Otwombe, Letutu, Kamariza, Sterling, Bertozzi, Martinson and Kana

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Participant disposition and enrollment microbiology. (A) Liquid limiting dilution assays (LLDs) to detect distinct differentially culturable populations in sputum samples. (B) Participant disposition flow chart. A total of 175 potential participants were screened for this study from primary health care clinics in the Soweto and Matlosana regions. From each cohort, the arrows depict participants who were removed from the study for various clinical and laboratory reasons. ^†Screen failures (18 from Soweto, 8 from Matlosana) include potential participants who were initially suitable for the study and then found to not meet the requirements. ^‡Participants were terminated from the study (21 from Soweto, 3 from Matlosana) and reasons for their removal are documented in the supplementary information. Methods optimization denoted by (^§) include LLDs performed for sputum samples with different plates and plate-sealing methods to address emerging evaporation of cultures and contamination issues. This aspect is further discussed in the supplementary material. ^¥Loss to follow up and one participant was excluded as no sputum sample was received for baseline LLD and CFU data. ^# One participant missed the day 180 time point but returned thereafter, the individual was excluded from the post 180 day analysis. Data from a total of 80 participants were available for baseline analysis. Additional participants were lost to follow up at various time points throughout the study. Those lost before or after day 35 were classified as early and late dropouts respectively. Data from 73 participants were available for early treatment analysis (sputum samples received up to day 14 following treatment initiation) and data from 62 participants were available for end of treatment analysis (sputum samples received up to 180 days following treatment initiation). (C) Bacterial yield using CF-supplemented LLDs (pink), Rpf ^- CF-supplemented LLDs (orange), un-supplemented LLDs (green) and colony forming units (purple) in enrollment sputum from TB and TB-HIV infected individuals. *p<0.05, **p<0.01, ****p<0.0001. ns = Not significant.