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High Mycobacterium tuberculosis bacillary loads detected by tuberculosis molecular bacterial load assay in patient stool : a potential alternative for nonsputum diagnosis and treatment response monitoring of tuberculosis

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Date
23/02/2022
Author
Musisi, Emmanuel
Sessolo, Abdul
Kaswabuli, Sylvia
Zawedde, Josephine
Byanyima, Patrick
Kasiinga, Shariifah
Sanyu, Ingvar
Uwimaana, Esther
Walimbwa, Stanley
Ola, Joseph
Ssengooba, Willy
Sekaggya, Christine
Joloba, Moses L.
Worodria, William
Huang, Laurence
Gillespie, Stephen Henry
Sloan, Derek James
Sabiiti, Wilber
Funder
Scottish Funding Council
Grant ID
N/A
Keywords
Molecular bacterial load assay
Molecular diagnostics
Mycobacterium tuberculosis
QR Microbiology
NDAS
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Abstract
Not all patients produce sputum, yet most available TB tests use sputum. We investigated the utility of a novel RNA-based quantitative test, the tuberculosis molecular bacterial load assay (TB-MBLA), for the detection and quantification of Mycobacterium tuberculosis in stool. Stools from 100 adult individuals were treated with OMNIgene-sputum reagent and tested using Xpert MTB/RIF ultra (Xpert ultra), auramine O smear microscopy (smear), mycobacterial growth indicator tube (MGIT), and Lowenstein-Jensen (LJ) cultures. The remaining portions were frozen at −20°C and later tested by TB-MBLA. MGIT sputum culture was used as a TB confirmatory test and reference for stool tests. Sixty-one of 100 participants were already confirmed TB positive by MGIT sputum culture, 20 (33%) of whom were HIV coinfected. TB-MBLA detected M. tuberculosis in 57/100 stool samples, including 49 already confirmed for TB. The mean bacterial load measured by stool TB-MBLA was 5.67 ± 1.7 log10 estimated CFU (eCFU) per mL in HIV-coinfected participants, which was higher than the 4.83 ± 1.59 log10 eCFU per mL among the HIV-negative participants (P = 0.04). The sensitivities (95% confidence intervals [CI]) of stool assays were 80% (68 to 89) and 90% (79 to 98) for TB-MBLA and Xpert ultra, which were both higher than the 44% (32 to 58), 64% (51 to 76), and 62% (45 to 77) for smear, MGIT, and Lowenstein-Jensen (LJ) stool cultures, respectively. The specificity (95% CI) of stool assays was highest for smear, at 97% (87 to 100), followed by Xpert ultra at 91% (76 to 98), TB-MBLA at 79% (63 to 90), LJ at 80% (64 to 91), and MGIT at 62% (45 to 77). Twenty-six percent of MGIT and 21% of LJ stool cultures were indeterminate due to contamination. Detection and quantification of viable M. tuberculosis bacilli in stool raises its utility as an alternative to sputum as a sample type for TB diagnosis.
Citation
Musisi , E , Sessolo , A , Kaswabuli , S , Zawedde , J , Byanyima , P , Kasiinga , S , Sanyu , I , Uwimaana , E , Walimbwa , S , Ola , J , Ssengooba , W , Sekaggya , C , Joloba , M L , Worodria , W , Huang , L , Gillespie , S H , Sloan , D J & Sabiiti , W 2022 , ' High Mycobacterium tuberculosis bacillary loads detected by tuberculosis molecular bacterial load assay in patient stool : a potential alternative for nonsputum diagnosis and treatment response monitoring of tuberculosis ' , Microbiology Spectrum , vol. 10 , no. 1 , e02100-21 . https://doi.org/10.1128/spectrum.02100-21
Publication
Microbiology Spectrum
Status
Peer reviewed
DOI
https://doi.org/10.1128/spectrum.02100-21
ISSN
2165-0497
Type
Journal article
Rights
Copyright © 2022 Musisi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
Description
Funding: Emmanuel’s doctoral research is supported by European and Developing countries Clinical Trial Partnership (EDCTP)-funded PanACEA II studentship (TR1A2015-1102), and the University of St Andrews St Leonards scholarship. Funding from the Infectious Diseases Institute, Makerere University to Mr Emmanuel Musisi and Dr Abdul Sessolo through Health and Innovation Impact project supported collection of specimens. Enrolment was funded by the Lung MicroCHIP (NIH: U01 HL098964) and K24 (NIH: K24 HL087713) grants through Professor Laurence Huang. Funding from the Scottish Funding Council (SCF)-Global401 Challenges Research Fund (GCRF) supported the TB-MBLA processing of the samples.
Collections
  • University of St Andrews Research
URI
http://hdl.handle.net/10023/24655

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