A prospective multicentre study was conducted to compare the diagnostic accuracy of the GeneXpert MTB/RIF Ultra (Xpert Ultra) and cell-free DNA (cfDNA) assay for tuberculous meningitis (TBM) in China. A prospective cohort study was conducted among individuals with symptoms suggestive of TBM registered in three TB specialised hospitals in China between June 2018 and January 2019. Overall, 84 patients suggestive of TBM were included in this analysis between June 2018 and January 2019. Using microbiological evidence as reference, the sensitivity/specificity for the diagnostic tests were Xpert Ultra 93.3%/100%, cfDNA 93.3%/92.6% and mycobacteria growth indicator tube (MGIT) culture 13.3%/100%. In addition, the sensitivity of culture was 6.7% when using clinical diagnosis criteria as the gold standard. Xpert Ultra correctly identified 28 cases as TBM, indicating a sensitivity of 46.7%. Notably, four additional TBM cases were detected by cfDNA compared with Xpert Ultra, yielding an overall sensitivity of 53.3%. Statistical analysis revealed that the sensitivity of Xpert Ultra and cfDNA was significantly higher than that of culture. The data demonstrate that Xpert Ultra and cfDNA assay showed optimal sensitivity compared with MGIT culture. In addition, there was no significant correlation between bacterial load and TBM severity in the participants.

Purpose of this paper is to analyze different species’ proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA,rpoBandhsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates’erm(41)ofM. abscessuswere sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged toM. aviumcomplex (MAC), and 70 isolates (25%) belonged toM. abscessus. Nearly all theM. abscessussubsp.abscessusresistant to clarithromycin had T28 inerm(41). However, all theM. abscessussubsp.abscessussusceptible to clarithromycin had C28 inerm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one wasM. abscessus. However,M. chelonei, M. fuerth, andM. gordonwere rare. Clarithromycin had a good inhibition activity against all the NTM species exceptM. abscessussubsp.abscessus. Theerm(41)genotype is of high relevance to clarithromycin resistance.

Objective: To investigate the in vitro activities of five oxazolidinones in parallel against the reference strains of different mycobacterial species and clinical isolates of Mycobacterium tuberculosis (Mtb), and shed light on the differences in the efficacy of these homolog drugs.Materials and methods: The minimum inhibitory concentrations (MICs) of linezolid, tedizolid, sutezolid, delpazolid, and contezolid against 16 mycobacterial reference strains and 69 M. tuberculosis clinical isolates, including 17 drug-susceptible isolates and 52 multidrug-resistant (MDR) isolates, were determined by microplate alamarBlue assay (MABA). The intracellular killing activities of contezolid and linezolid against Mtb H37Rv were compared. In addition, mutations in the linezolid resistance-related genes (rplC, rplD, and 23S rRNA) of the Mtb clinical isolates were also analyzed.Results: Tedizolid exhibited the strongest inhibitory activities against the reference strains of both rapidly growing mycobacteria (RGM) and slowly growing mycobacteria (SGM), among the tested oxazolidinones. In contrast, sutezolid only manifested potent activity against reference strains of SGM. Linezolid, delpazolid, and contezolid were less active against the non-tuberculous mycobacterial references. For the Mtb clinical isolates, the antimicrobial action was ranked as: sutezolid > tedizolid > contezolid and linezolid > delpazolid, whereas no difference between drug-sensitive and multiple drug-resistant isolates was observed. Notably, contezolid demonstrated obviously superior intracellular antimicrobial activity than linezolid. Few strains harbored mutations in rrl gene or rplD genes, although these strains had drug susceptible profiles to linezolid.Conclusion: Different oxazolidinones can have discrepant antimicrobial activity against different mycobacterial species, or have different manifestations out of cell or in cell. Understanding these differences would be helpful in choosing the appropriate drug in clinical practice.

Background. Treatment response for the Mycobacterium abscessus (M. abscessus) lung disease remains far from satisfying. An effective regimen is needed to solve the problem. Methods. We retrospectively reviewed the medical records of all patients with M. abscessus lung disease who received antibiotics regimen at Beijing Chest Hospital Affiliated to Capital Medical University between July 1, 2010, and February 1, 2018. Patients were administered a conventional antibiotics regimen (including macrolide and moxifloxacin, along with an initial 12-week course of low-dose cefoxitin and amikacin) or intensified regimen (including a higher dosage of cefoxitin and linezolid besides conventional drugs), respectively. The time to sputum-culture conversion and proportion of sputum-culture conversion in liquid broth were investigated to evaluate the efficacy and evaluation of safety by performing the classification of adverse events according to the Division of AIDS, National Institute of Allergy and Infectious Disease. Patients were followed for 18 months from baseline. Results. In the conventional regimen group, the sputum conversion rate at 18 months was 29.4% (10/34), and the median time until sputum conversion was 2 months (IQR, 1-2 mo). Furthermore, in the intensified regimen group, the sputum conversion rate was 81.3% (13/16), and the median time until sputum conversion was 1 month (IQR, 1-1 mo). Leukopenia and drug-induced hepatotoxicity occurred more frequently in the intensified regimen group in contrast with the conventional regimen group patients. However, only 1 adverse event in the intensified regimen group was classified as severe adverse event. Conclusions. The intensified regimen could improve sputum conversion of M. abscessus lung disease compared with conventional regimen, but close safety surveillance is necessary to monitor adverse events.

Objectives. To evaluate the resolution and reliability of therpsAgene, encoding ribosomal protein S1, as a novel biomarker for mycobacteria species identification.Methods. A segment of therpsAgene (565 bp) was amplified by PCR from 42 mycobacterial reference strains, 172 nontuberculosis mycobacteria clinical isolates, and 16M. tuberculosiscomplex clinical isolates. The PCR products were sequenced and aligned by using the multiple alignment algorithm in the MegAlign package (DNASTAR) and the MEGA program. A phylogenetic tree was constructed by the neighbor-joining method.Results. Comparative sequence analysis of therpsAgene provided the basis for species differentiation within the genusMycobacterium. Slow- and rapid-growing groups of mycobacteria were clearly separated, and each mycobacterial species was differentiated as a distinct entity in the phylogenetic tree. The sequences discrepancy was obvious betweenM. kansasiiandM. gastri, M. chelonaeandM. abscessus, M. aviumandM. intracellulare, andM. szulgaiandM. malmoense, which cannot be achieved by 16S ribosomal DNA (rDNA) homologue genes comparison. 183 of the 188 (97.3%) clinical isolates, consisting of 8 mycobacterial species, were identified correctly byrpsAgene blast.Conclusions. Our study indicates thatrpsAsequencing can be used effectively for mycobacteria species identification as a supplement to 16S rDNA sequence analysis.