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Evaluation of the Analytical Performance of the Xpert MTB/RIF Assay
Open accessAbstract
We performed the first studies of analytic sensitivity, analytic specificity, and dynamic range for the new Xpert MTB/RIF assay, a nucleic acid amplification-based diagnostic system that detects Mycobacterium tuberculosis and rifampin (RIF) resistance in under 2 h. The sensitivity of the assay was tested with 79 phylogenetically and geographically diverse M. tuberculosis isolates, including 42 drug-susceptible isolates and 37 RIF-resistant isolates containing 13 different rpoB mutations or mutation combinations. The specificity of the assay was tested with 89 nontuberculosis bacteria, fungi, and viruses. The Xpert MTB/RIF assay correctly identified all 79 M. tuberculosis isolates and correctly excluded all 89 nontuberculosis isolates. RIF resistance was correctly identified in all 37 resistant isolates and in none of the 42 susceptible isolates. Dynamic range was assessed by adding 10(2) to 10(7) CFU of M. tuberculosis into M. tuberculosis-negative sputum samples. The assay showed a log-linear relationship between cycle threshold and input CFU over the entire concentration range. Resistance detection in the presence of different mixtures of RIF-resistant and RIF-susceptible DNA was assessed. Resistance detection was dependent on the particular mutation and required between 65% and 100% mutant DNA to be present in the sample for 95% certainty of resistance detection. Finally, we studied whether assay specificity could be affected by cross-contaminating amplicons generated by the GenoType MTBDRplus assay. M. tuberculosis was not detected until at least 10(8) copies of an MTBDRplus amplicon were spiked into 1 ml of sputum, suggesting that false-positive results would be unlikely to occur.
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“Previous studies have shown that Xpert does not cross‐react with other mycobacterial species (Blakemore 2010; Helb 2010).”
From Xpert MTB/RIF Ultra and Xpert MTB/RIF assays for extrapulmonary tuberculosis and rifampicin resistance in adults (2021) · cites this paper as Blakemore 2010
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Rapid Molecular Detection of Tuberculosis and Rifampin Resistance
Boehme, C. C., Nabeta, P., Hillemann, D., Nicol, M. P., Shenai, S., Krapp, F., Allen, J., Tahirli, R., Blakemore, R., Rustomjee, R., et al. (2010) · New England Journal of Medicine · doi:10.1056/nejmoa0907847 · Cited by 2,309
Self-citationXpert® MTB/RIF assay for pulmonary tuberculosis and rifampicin resistance in adults
Steingart, K. R., Schiller, I., Horne, D. J. D. L., Pai, M., Boehme, C. C., & Dendukuri, N. (2014) · Cochrane Database of Systematic Reviews · doi:10.1002/14651858.cd009593.pub3 · Cited by 1,187
Point-of-care nucleic acid testing for infectious diseases
Niemz, A., Ferguson, T. M., & Boyle, D. S. (2011) · Trends in biotechnology · doi:10.1016/j.tibtech.2011.01.007 · Cited by 838
Chakravorty, S., Simmons, A. M., Rowneki, M., Parmar, H., Cao, Y., Ryan, J., Banada, P. P., Deshpande, S., Shenai, S., Gall, A., et al. (2017) · mBio · doi:10.1128/mbio.00812-17 · Cited by 668
Ellington, M. J., Ekelund, O., Aarestrup, F. M., Canton, R., Doumith, M., Giske, C., Grundman, H., Hasman, H., Holden, M. T. G., Hopkins, K. L., et al. (2017) · Clinical Microbiology and Infection · doi:10.1016/j.cmi.2016.11.012 · Cited by 603
Metadata
- Authors
- Blakemore, R., Story, E., Helb, D., Kop, J., Banada, P., Owens, M. R., Chakravorty, S., Jones, M., & Alland, D.
- Affiliations
- Rutgers New Jersey Medical School (US)
- Division of Infectious Disease, Department of Medicine, and Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey
- Cepheid (United States) (US)
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2010-07
- Volume
- 48
- Issue
- 7
- Pages
- 2495-2501
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC2897495
- OpenAlex
- W2132658179
- Cited by
- 485
- References
- 21
- Citations by year
- 2026: 4 · 2025: 14 · 2024: 19 · 2023: 18 · 2022: 27 · 2021: 31 · 2020: 32 · 2019: 34 · 2018: 32 · 2017: 52 · 2016: 30 · 2015: 36 · 2014: 39 · 2013: 32 · 2012: 28
- FWCI
- 24.27
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Clinical Microbiology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Rutgers University Community Repository (Rutgers University) (submittedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (R41AI052523, R42AI052523, R42-AI52523, R41-AI52523)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Health Sciences › Medicine › Infectious Diseases › Infectious Diseases and Tuberculosis
- Keywords
- Xpert MTB/RIF, Mycobacterium tuberculosis, rifampicin resistance, rpoB mutations, diagnostic sensitivity, diagnostic specificity, dynamic range, cross-contamination, GenoType MTBDRplus, sputum
- MeSH terms
- Antibiotics, Antitubercular, Bacterial Proteins, DNA, Bacterial, Microbial Sensitivity Tests, Mycobacterium tuberculosis, Rifampin, DNA-Directed RNA Polymerases, Sensitivity and Specificity, Sputum, Reproducibility of Results, Linear Models, Nucleic Acid Amplification Techniques, Molecular Diagnostic Techniques
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