4.2 Article

Supplementation of sputum cultures with culture filtrate to detect tuberculosis in a cross-sectional study of HIV-infected individuals

Journal

TUBERCULOSIS
Volume 129, Issue -, Pages -

Publisher

CHURCHILL LIVINGSTONE
DOI: 10.1016/j.tube.2021.102103

Keywords

Mycobacterium tuberculosis, MGIT time To positivity; Resuscitation promoting factors; Culture filtrate; Differentially culturable tubercle bacteria

Funding

  1. International Early Career Scientist Award from the Howard Hughes Medical Institute
  2. South African National Research Foundation
  3. South African Medical Research Council
  4. National Health Laboratory Services Research Trust
  5. National Health Scholars Programme
  6. Soweto Matlosana Collaborating Centre for HIV/Aids and Tuberculosis

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The addition of growth factors to the MGIT culture system can reduce the time to culture positivity, especially in HIV-TB co-infected patients with low sputum bacillary loads. This method has shown to enhance detection of M. tuberculosis in specimens that were previously negative under standard conditions.
While some healthcare systems have shifted to molecular diagnostics, culture still remains the gold standard for tuberculosis diagnosis, but it is limited by its long duration to a positive result. Methods to reduce time to culture positivity (TTP) are urgently required. We determined if growth factor supplementation in the mycobacterial growth indicator tube (MGIT) culture system reduces TTP. MGITs were supplemented with fresh culture filtrate (CF) as a source of growth stimulatory molecules from axenic Mycobacterium tuberculosis culture. Different volumes of CF and media components were tested. The performance of these modified MGITs was assessed with sputum from HIV-TB co-infected individuals. Reducing the volume of MGIT cultures and removal of detergent from cultures grown to generate CF had a marginal but significant benefit on reducing TTP. In a subset of specimens, CF inhibited growth. Following optimization of methods, a reduced TTP occurred in specimens with low bacillary load as measured by GeneXpert, smear microscopy and colony forming units. Three specimens that were negative under standard conditions flagged positive following CF supplementation. Our data provide preliminary evidence that addition of CF to MGIT cultures can enhance detection of M. tuberculosis in HIV-TB coinfected patients with low sputum bacillary loads.

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