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A luciferase-based ...
A luciferase-based assay for rapid assessment of drug activity against Mycobacterium tuberculosis including monitoring of macrophage viability
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- Larsson, Marie C (författare)
- Östergötlands Läns Landsting,Linköpings universitet,Avdelningen för mikrobiologi och molekylär medicin,Hälsouniversitetet,Klinisk mikrobiologi
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- Lerm, Maria (författare)
- Linköpings universitet,Avdelningen för mikrobiologi och molekylär medicin,Hälsouniversitetet
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- Angeby, Kristian (författare)
- Karolinska Institutet,Karolinska University Hospital, Hospital, Stockholm, Sweden; University of the West Indies, Kingston, Jamaica
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- Nordvall, Michaela (författare)
- Östergötlands Läns Landsting,Klinisk mikrobiologi
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- Jureen, Pontus (författare)
- Public Health Agency of Sweden, Stockholm, Sweden
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- Schön, Thomas (författare)
- Linköpings universitet,Linnéuniversitetet,Institutionen för medicin och optometri (MEO),Linköping Univ ; Kalmar County Hospital,Medicinsk mikrobiologi,Hälsouniversitetet,Kalmar County Hospital, Sweden; Linnaeus University, Kalmar, Sweden
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(creator_code:org_t)
- Elsevier BV, 2014
- 2014
- Engelska.
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Ingår i: Journal of Microbiological Methods. - : Elsevier BV. - 0167-7012 .- 1872-8359. ; 106, s. 146-150
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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http://kipublication...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- The intracellular (IC) effect of drugs against Mycobacterium tuberculosis (Mtb) is not well established but increasingly important to consider when combining current and future multidrug regimens into the best possible treatment strategies. For this purpose, we developed an IC model based on a genetically modified Mtb H37Rv strain, expressing the Vibrio harvei luciferase (H37Rv-lux) infecting the human macrophage like cell line THP-1. Cells were infected at a low multiplicity of infection (1:1) and subsequently exposed to isoniazid (INH), ethambutol (EMB), amikacin (AMI) or levofloxacin (LEV) for 5 days in a 96-well format. Cell viability was evaluated by Calcein AM and was maintained throughout the experiment. The number of viable H37Rv-lux was determined by luminescence and verified by a colony forming unit analysis. The results were compared to the effects of the same drugs in broth cultures. AMI, EMB and LEV were significantly less effective intracellularly (MIC90: >4 mg/L, 8 mg/L and 2 mg/L, respectively) compared to extracellularly (MIC90: 0.5 mg/L for AMI and EMB; 0.25 mg/L for LEV). The reverse was the case for INH (IC: 0.064 mg/L vs EC: 0.25 mg/L). In conclusion, this luciferase based method, in which monitoring of cell viability is included, has the potential to become a useful tool while evaluating the intracellular effects of anti-mycobacterial drugs. (C) 2014 Elsevier B.V. All rights reserved.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine (hsv//eng)
Nyckelord
- Tuberculosis
- Levofloxacin
- Drug susceptibility testing
- Intracellular
- Luminescence
- Minimal inhibitory concentration
- Biomedicinsk vetenskap
- Biomedical Sciences
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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