التفاصيل البيبلوغرافية
| العنوان: |
A novel mycobacterial growth inhibition assay employing live-cell imaging of virulent M. tuberculosis and monitoring of host cell viability |
| المؤلفون: |
Amanda Welin, Thomas B. Schön, Michaela Jonsson Nordvall, Maria Lerm, Blanka Andersson |
| المصدر: |
Tuberculosis. 124:101977 |
| بيانات النشر: |
Elsevier BV, 2020. |
| سنة النشر: |
2020 |
| مصطلحات موضوعية: |
0301 basic medicine, Microbiology (medical), Time Factors, Cell Survival, Green Fluorescent Proteins, 030106 microbiology, Immunology, Virulence, Microbiology, Peripheral blood mononuclear cell, Mycobacterium tuberculosis, 03 medical and health sciences, chemistry.chemical_compound, Multiplicity of infection, Genes, Reporter, Predictive Value of Tests, Live cell imaging, Humans, Tuberculosis, Viability assay, biology, Reproducibility of Results, biology.organism_classification, 030104 developmental biology, Infectious Diseases, Microscopy, Fluorescence, chemistry, Relative fluorescence units, Host-Pathogen Interactions, Leukocytes, Mononuclear, Growth inhibition |
| الوصف: |
Our aim was to develop a Mycobacterium tuberculosis (Mtb) growth inhibition assay (MGIA) as a summary estimate of host immune control of virulent Mtb. Mycobacterial growth inhibition (MGI) using previously frozen human PBMCs infected with H37Rv was assessed by live-cell imaging (Incucyte©) complemented by imaging flow cytometry analysis of phagocytosis. MGI measured as relative fluorescence units (RFU) was calibrated to time to positive culture (TTP) in BACTEC 960 MGIT. At a MOI (multiplicity of infection) of 5, there was a wide range of MGI of blood donors (1.1*106–2.7*106 RFU, n = 14). Intra- and inter-assay variability were at most 17.5 and 20.7 CV%. Cell viability at day 5 was 57 and 62% monitored by the LDH and Draq7 assays respectively. There was a strong correlation between a readout for Mtb growth using CFU counts or TTP compared to RFU (r2≥0.96). Our MGIA enabling live-cell imaging and monitoring of cell viability was able to detect a wide range of Mtb growth inhibition by PBMCs and was calibrated to several readout options for bacterial growth. This MGIA may be valuable as a surrogate marker of host immunity in a personalized medicine approach. |
| تدمد: |
1472-9792 |
| الوصول الحر: |
https://explore.openaire.eu/search/publication?articleId=doi_dedup___::542ec5799cf31ffd0cf83e660da4f4d5Test
https://doi.org/10.1016/j.tube.2020.101977Test |
| حقوق: |
CLOSED |
| رقم الانضمام: |
edsair.doi.dedup.....542ec5799cf31ffd0cf83e660da4f4d5 |
| قاعدة البيانات: |
OpenAIRE |
