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1.  Evaluation of Cobas TaqMan MTB for Direct Detection of the Mycobacterium tuberculosis Complex in Comparison with Cobas Amplicor MTB 
Journal of Clinical Microbiology  2013;51(7):2112-2117.
The Roche Cobas Amplicor MTB assay, recently replaced by the Roche Cobas TaqMan MTB assay, was one of the first commercially available assays for detection of the Mycobacterium tuberculosis complex based on nucleic acid amplification. We reported previously on the limited specificity of the Cobas Amplicor MTB assay, in particular for positive samples with an optical density at 660 nm (OD660) of <2.0. Using a selected set of respiratory samples, which were scored as false positive by the Cobas Amplicor test, we demonstrate here that the specificity of the Cobas TaqMan assay is significantly improved. In addition, our study of a set of 133 clinical samples revealed that the Cobas TaqMan MTB assay showed significantly less PCR inhibition than the Cobas Amplicor test. An overall concordance of 98.2% was observed between the two assays. In a subsequent prospective study, we evaluated the performance of the Roche Cobas TaqMan MTB assay on 1,143 clinical specimens, including respiratory (n = 838) and nonrespiratory (n = 305) specimens. Using culture as the gold standard, we found a sensitivity of 88.4% and a specificity of 98.8% for the 838 respiratory specimens, compared to a sensitivity of 63.6% and a specificity of 94.6% for the 305 nonrespiratory specimens. We conclude that the Cobas TaqMan MTB assay is a significantly improved tool for the direct detection of M. tuberculosis DNA in clinical specimens.
doi:10.1128/JCM.00142-13
PMCID: PMC3697670  PMID: 23616457
2.  Septicemia Caused by Tick-borne Bacterial Pathogen Candidatus Neoehrlichia mikurensis 
Emerging Infectious Diseases  2010;16(7):1127-1129.
We have repeatedly detected Candidatus Neoehrlichia mikurensis, a bacterium first described in Rattus norvegicus rats and Ixodes ovatus ticks in Japan in 2004 in the blood of a 61-year-old man with signs of septicemia by 16S rRNA and groEL gene PCR. After 6 weeks of therapy with doxycycline and rifampin, the patient recovered.
doi:10.3201/eid1607.091907
PMCID: PMC3358111  PMID: 20587186
Candidatus Neoehrlichia mikurensis; septicemia; human infection; 16S rRNA gene PCR; therapy; tick-borne pathogen; bacteria; dispatch
3.  Quantitative Drug Susceptibility Testing of Mycobacterium tuberculosis by Use of MGIT 960 and EpiCenter Instrumentation ▿  
Journal of Clinical Microbiology  2009;47(6):1773-1780.
Since numbers of drug-resistant Mycobacterium tuberculosis strains are on the rise, the simple classification into “susceptible” and “resistant” strains based on susceptibility testing at “critical concentrations” has to be reconsidered. While future studies have to address the correlation of phenotypic resistance levels and treatment outcomes, a prerequisite for corresponding investigations is the ability to exactly determine levels of quantitative drug resistance in clinical M. tuberculosis isolates. Here we have established the conditions for quantitative drug susceptibility testing for first- and second-line agents using MGIT 960 instrumentation and EpiCenter software equipped with the TB eXiST module. In-depth comparative analysis of a range of well-characterized susceptible and resistant clinical isolates has allowed us to propose conditions for testing and to develop criteria for interpretation.
doi:10.1128/JCM.02501-08
PMCID: PMC2691107  PMID: 19339475
4.  Tuberculosis Drug Resistance in an Area of Low Endemicity in 2004 to 2006: Semiquantitative Drug Susceptibility Testing and Genotyping▿  
Journal of Clinical Microbiology  2008;46(12):4064-4067.
We determined the quantitative levels and the genetic mechanisms of resistance in drug-resistant clinical isolates of Mycobacterium tuberculosis sampled over a period of 3 years (n = 45; 17 of the isolate were multidrug resistant). Our results led us to hypothesize that some strains categorized as resistant to isoniazid, ethambutol, or streptomycin by standard laboratory procedures of in vitro drug susceptibility testing may still respond to a treatment regimen that includes these agents.
doi:10.1128/JCM.01114-08
PMCID: PMC2593253  PMID: 18923010

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