Quantification of viable bacterial load in artificial sputum spiked with Mycobacterium tuberculosis
| Autor: | Lize van der Merwe, Andreas H. Diacon, Sven O. Friedrich, Miriam N. Karinja, Eva Kolwijck |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Sample (material) 030106 microbiology Immunology Microbiology Specimen Handling Mycobacterium tuberculosis 03 medical and health sciences All institutes and research themes of the Radboud University Medical Center medicine Time to positivity Chromatography biology Sputum Reproducibility of Results biology.organism_classification Bacterial Load Culture Media lnfectious Diseases and Global Health Radboud Institute for Health Sciences [Radboudumc 4] 030104 developmental biology Infectious Diseases Mycobacterium tuberculosis H37Rv Feasibility Studies Variance components High load medicine.symptom Stock solution |
| Zdroj: | Tuberculosis, 115, 140-145 Tuberculosis, 115, pp. 140-145 |
| ISSN: | 1472-9792 |
| Popis: | Objective Artificial sputum spiked with Mycobacterium tuberculosis could serve for validation of procedures that determine viable mycobacterial load. Design Artificial sputum specimens prepared in-house were spiked with low, medium or high concentrations of Mycobacterium tuberculosis H37Rv stock solution. In a first series, a single technologist processed two batches of specimens daily with high load that were stored refrigerated or at room temperature for up to 8 days. In a second series, nine different technologists processed freshly made batches of specimens with low, medium or high loads. We recorded time to positivity (TTP) in duplicate liquid cultures made from each specimen. Results Specimens were well grouped around the mean TTP (hours; standard deviation) of low: 271.7 (25.9), medium: 233.5 (16.3), and two batches of high load: 186.9 (12.3) and 191.8 (9.0), respectively. A variance component model that included load, storage temperature, days of storage until processing, batch of specimens made, sample ID and technologist ID as random effects in a linear mixed-effects model identified only load, technologist and residual as significant contributors to overall TTP variance. Conclusion Artificial sputum specimens with reproducible and stable viable mycobacterial loads can be made that could serve for training and validation purposes. |
| Databáze: | OpenAIRE |
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