Affiliation:
1. DST/NRF Centre of Excellence for Biomedical TB Research, School of Pathology, Faculty of Health Sciences, University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa
2. MRC-CAPRISA HIV-TB Pathogenesis and Treatment Research Unit, Centre for the AIDS Programme of Research in South Africa, Durban, South Africa
Abstract
ABSTRACT
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.
Funder
National Research Foundation of South Africa
Wits Enterprize
Technology Innovation Agency
Howard Hughes Medical Institute
South African Medical Research Council
Department of Science and Technology, Republic of South Africa
Publisher
American Society for Microbiology
Cited by
6 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献