Metagenomic next-generation sequencing (mNGS) is an effective diagnostic method for infectious diseases, however, its clinical utility for tuberculosis (TB) diagnosis remains to be demonstrated. A total of 322 bronchoalveolar lavage fluid (BALF) samples were collected from 311 suspected and confirmed pulmonary TB patients and tested by mNGS, acid-fast bacillus (AFB) smear by microscopy, Xpert® MTB/RIF (Xpert), mycobacterium culture and bacterial/fungal culture. Diagnostic performance of mNGS was compared with conventional methods for detection of Mycobacterium tuberculosis complex (MTBC) and other pathogens in BALF. Underlying factors associated with positive detection in pulmonary TB patients were investigated. mNGS, Xpert and culture presented a high proportion of complete matching for MTBC detection (244/322, 75.8%). In pulmonary TB patients pre-treatment the sensitivity of MTBC detection by mNGS, Xpert, culture and smear was 59.9% (85/142), 69.0% (98/142), 59.9% (85/142) and 24.6% (35/142), respectively, and 79.6% overall; MTBC was detected by mNGS in 33.2% (5/34) Xpert and culture negative samples. Positive MTBC detection by mNGS was affected by Vitamin D, erythrocyte sedimentation rate, TB initial treatment/retreatment, and cavity in chest imaging (χ2 = 37.42, P < 0.001), but not by prior anti-TB therapy within 3 months. mNGS was able to detect new potential pathogens in 8.7% (28/322) of samples. Combining mNGS with conventional detection methods could increase the detection rate for MTBC. Additionally, mNGS could identify pathogens in a non-targeted approach for better diagnosis of coinfection.

Histoplasmosis is an endemic disease mainly occurring in North America and is rare in China. Confirmation of histoplasmosis should be based on a compatible clinical scenario and a positive culture or histopathology. However, there are still many cases that are misdiagnosed or missed, especially in individuals from non-endemic areas. In recent years, metagenomic next-generation sequencing (mNGS) has led to the accurate and timely diagnosis of some rare and complicated infectious diseases. We describe the case of a 27-year-old Chinese man who had chronic progressive pulmonary lesions without any symptoms for more than 1year, until the lesions reached the epiglottis and led to progressive pharyngeal pain. There were no positive results from bronchoalveolar lavage fluid (BALF) and epiglottis tissue cultures, or from epiglottis and lung pathological examinations, but mNGS was able to identify Histoplasma capsulatum in the epiglottis tissues and BALF as the cause of the lesions. The patient was treated successfully with itraconazole.

Background: Moxifloxacin (MFX, M) is currently a second-line antituberculosis drug as initial therapy of pulmonary tuberculosis and one of the main anti-TB drugs in drug-resistant TB, which can kill both intracellular and extracellular Mycobacterium tuberculosis. We started a trial to study the efficacy and safety of the substitution of pyrazinamide and ethambutol with moxifloxacin during the intensive phase of treatment of newly diagnosed susceptive pulmonary tuberculosis. Methods/design: This is a prospective, open, randomized, parallel-controlled, single-center clinical study. The study consists of three phases: a screening period, a treatment period of 6 (or 7) months, and a follow-up period of 1 year. Patients selected for the study will be allocated to the trial group or the control group randomly. The control group will be given six months of a standard regimen(2HRZE/4HR). The trial group will be given a total of six months of treatment with substitution of pyrazinamide and ethambutol with moxifloxacin during the intensive phase(2HRM/4HR). The primary outcome is the rate of adverse outcomes within one year of completion of therapy. The Secondary outcomes include the rate of treatment success at the 2nd, 3rd, 5th and 6th months, the rate of sputum Mtb(Mycobacterium tuberculosis) negative conversion at the 2nd, 3rd, 5th and 6th months, the time of sputum Mtb negative conversion at the 2nd, 3rd, 5th and 6th months, and the number of patients with adverse events within one year of completion of therapy. Comparisons will be performed using two-sided tests with a statistical significance level of 5%.Discussion: This trial will reveal the effectiveness and safety of 2months of use of moxifloxacin instead of pyrazinamide and ethambutol during the intensive phase of treatment for newly diagnosed susceptive pulmonary tuberculosis. If the new regimen including isoniazid, rifampicin and moxifloxacin during the intensive phase of treatment (2HRM/4HR) is no less effective and safe than the standard regimen(2HRZE/4HR), it could be a new alternative treatment for newly diagnosed susceptive pulmonary tuberculosis in the future. Trial registration: ClinicalTrials.gov, NCT04187469. Registered on 5 December 2019.