Natural polymorphisms in mycobacterium tuberculosis conferring resistance to delamanid in drug-naïve patients.

Mutations in the genes of the F420 signaling pathway, including dnn, fgd1, fbiA, fbiB, fbiC, and fbiD, of Mycobacterium tuberculosis (Mtb) complex can lead to delamanid resistance. We searched for such mutations among 129 Mtb strains from Asia, South-America, and Africa using whole-genome sequencing; 70 (54%) strains had at least one mutation in one of the genes. For ten strains with mutations, we determined the minimum inhibitory concentration (MIC) of delamanid. We found one strain from a delamanid-naïve patient carrying the natural polymorphism Tyr29del (ddn) that was associated with a critical MIC to delamanid

Identification of Eps15 as Antigen Recognized by the Monoclonal Antibodies aa2 and ab52 of the Wuerzburg Hybridoma Library against Drosophila Brain

The Wuerzburg Hybridoma Library against the Drosophila brain represents a collection of around 200 monoclonal antibodies that bind to specific structures in the Drosophila brain. Here we describe the immunohistochemical staining patterns, the Western blot signals of one- and two-dimensional electrophoretic separation, and the mass spectrometric characterization of the target protein candidates recognized by the monoclonal antibodies aa2 and ab52 from the library. Analysis of a mutant of a candidate gene identified the Drosophila homolog of the Epidermal growth factor receptor Pathway Substrate clone 15 (Eps15) as the antigen for these two antibodies

Mortality of drug-resistant tuberculosis in high-burden countries: comparison of routine drug susceptibility testing with whole-genome sequencing

Background: Drug-resistance threatens global tuberculosis control. We examined mortality in patients with tuberculosis from high-burden countries, according to concordance or discordance of results from drug susceptibility testing done locally and whole-genome sequencing (WGS). Methods: We collected pulmonary Mycobacterium tuberculosis isolates and clinical data from adult tuberculosis patients from Côte d'Ivoire, Democratic Republic of the Congo, Kenya, Nigeria, South Africa, Peru, and Thailand, stratified by HIV status and drug resistance, from 2013 to 2016. Sites tested drug susceptibility using routinely available methods. WGS was done on Illumina HiSeq 2500 in the USA and Switzerland, and TBprofiler used to analyse the genomes. We analysed mortality in multivariable logistic regression models adjusted for sex, age, HIV-status, history of tuberculosis, and sputum positivity. Findings: We included 582 tuberculosis patients. The median age was 33 years (interquartile range 27-43 years), 225 (39%) were female, and 247 (42%) were HIV-positive. Based on WGS, 339 (58%) isolates were pan- susceptible, 35 (6%) monoresistant, 146 (25%) multidrug-resistant, and 24 (4%) pre-/ extensively drug-resistant (pre-XDR/XDR). The local results were discordant with the WGS results in 130/582 (22%) of patients. All testing methods identified isoniazid and rifampicin resistance with a high agreement. Resistance to ethambutol, pyrazinamide, and second-line drugs was rarely tested locally. Of 576 patients with known treatment, 86 (15%) patients received inappropriate treatment according to WGS results and the World Health Organization (WHO) treatment guidelines. The analysis of mortality was based on 530 patients; 63 (12%) died, and 77 patients (15%) received inappropriate treatment. Mortality ranged from 6% in patients with pan-susceptible tuberculosis (18/310) to 39% in patients with pre-XDR/XDR tuberculosis (9/23). The adjusted odds ratio for mortality was 4.92 (95% CI 2.47-9.78) among under-treated patients, compared to appropriately treated cases. Interpretation: In seven high-burden tuberculosis countries, we observed discrepancies between drug resistance patterns obtained locally and WGS. The under-diagnosis of drug resistances resulted in inappropriate treatment and higher mortality. WGS can provide accurate and detailed drug resistance information required to improve the outcomes of drug-resistant tuberculosis in high-burden settings. Our results support the WHO's call for point-of- care tests based on WGS