Genetic Mimetics of Mycobacterium tuberculosis and Methicillin-Resistant Staphylococcus aureus as Verification Standards for Molecular Diagnostics
| Autor: | Edith E. Machowski, Bavesh D. Kana |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
DNA
Bacterial Methicillin-Resistant Staphylococcus aureus 0301 basic medicine Microbiology (medical) Tuberculosis Mycobacterium smegmatis 030106 microbiology medicine.disease_cause Microbiology Mycobacterium tuberculosis 03 medical and health sciences medicine Humans Pathology Molecular Recombination Genetic GeneXpert MTB/RIF biology SCCmec Mycobacteriology and Aerobic Actinomycetes Reference Standards Staphylococcal Infections Molecular diagnostics rpoB medicine.disease biology.organism_classification Methicillin-resistant Staphylococcus aureus Virology 030104 developmental biology Mutation |
| Zdroj: | Journal of Clinical Microbiology. 55:3384-3394 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.01111-17 |
| Popis: | Molecular diagnostics have revolutionized the management of health care through enhanced detection of disease or infection and effective enrollment into treatment. In recognition of this, the World Health Organization approved the rollout of nucleic acid amplification technologies for identification of Mycobacterium tuberculosis using platforms such as GeneXpert MTB/RIF, the GenoType MTBDR plus line probe assay, and, more recently, GeneXpert MTB/RIF Ultra. These assays can simultaneously detect tuberculosis infection and assess rifampin resistance. However, their widespread use in health systems requires verification and quality assurance programs. To enable development of these, we report the construction of genetically modified strains of Mycobacterium smegmatis that mimic the profile of Mycobacterium tuberculosis on both the GeneXpert MTB/RIF and the MTBDR plus line probe diagnostic tests. Using site-specific gene editing, we also created derivatives that faithfully mimic the diagnostic result of rifampin-resistant M. tuberculosis , with mutations at positions 513, 516, 526, 531, and 533 in the rifampin resistance-determining region of the rpoB gene. Next, we extended this approach to other diseases and demonstrated that a Staphylococcus aureus gene sequence can be introduced into M. smegmatis to generate a positive response for the SCC mec probe in the GeneXpert SA Nasal Complete molecular diagnostic cartridge, designed for identification of methicillin-resistant S. aureus . These biomimetic strains are cost-effective, have low biohazard content, accurately mimic drug resistance, and can be produced with relative ease, thus illustrating their potential for widespread use as verification standards for diagnosis of a variety of diseases. |
| Databáze: | OpenAIRE |
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