The purpose of this study was to evaluate the application value of targeted next-generation sequencing(tNGS)for rapid detection of Mycobacterium tuberculosis.From September 2021 to October 2024,1 178 clinical samples from hospitalized pa-tients at the Non-tuberculous Mycobacteria Disease Diagnosis and Treatment Center of Guangzhou Chest Hospital were collected,in-cluding 272 sputum samples,871 broncho alveolar lavage fluid(BALF)samples,and 35 other samples.These samples were analyzed with tNGS and the liquid culture+DNA microarray chip method(referred to as the"culture identification method"),and the detection results between methods were compared.The numbers of Mycobacterium tuberculosis complex(MTBC)samples identified with tNGS and the culture identification method were 172 and 127,respectively,and the numbers of non-tuberculous mycobacteria(NTM)samples detected were 517 and 474,respectively.The detection rate of Mycobacterium was 56.88%with tNGS and was significantly lower(49.49%)with the culture identification method(χ2=13.27,P<0.05).For NTM identification,when sputum was used,the detection rate of tNGS was higher than that of the culture identification method(χ2=24.14,P<0.05).However,when BALF was used,no significant difference in detection rates was observed between tNGS and the culture identification method(χ2=3.97,P=0.06).When different sample types from the same patient were analyzed with tNGS for NTM,BALF was found to be a better choice than sputum(χ2=4.05,P<0.05).However,with the culture identification method,we observed no significant difference between sample types(χ2=2.72,P=0.10).Furthermore,both the MTBC and NTM sequences detected with tNGS were significantly higher in the culture-positive group than the culture-negative group.With increasing sequence numbers,the proportion of cases in the culture-positive group progressively rose,whereas an inverse trend was observed in the culture-negative group.With clinical diagno-sis as the reference standard,the sensitivity,specificity,and Kappa value of tNGS and the culture identification method in identifying mycobacterial diseases were 85.41%,91.32%,0.74 and 74.73%,93.15%,0.63,respectively.Compared with the culture identifica-tion method,tNGS for rapid detection of Mycobacterium exhibited excellent sensitivity,specificity and consistency,and its timeliness should help clinicians make precise individualized treatment plans earlier.

In recent year，the iatrogenic infections and outbreaks caused by Mycobacterium abscessus complex（MABC）have been increasingly reported. Studies indicate that MABC accounts for 65%-80% of rapidly growing non-tuberculosis Mycobacterium（NTM）-related pulmonary diseases. As a clinically significant NTM pathogen，MABC exhibits rapid proliferation and intrinsic resistance to multiple antimicrobial agents. Notably，the 16S rDNA sequences among MABC subspecies share 100% homology，posing challenges for subspecies-level differentiation. Rapid and precise identification of MABC at the species or subspecies level is therefore critical for effective clinical management and outbreak control. Conventional biochemical methods for NTM identification are time-consuming and prone to inaccuracies，whereas molecular biology techniques offer superior speed and specificity，and have been widely used clinically. This review highlights the clinical implications of molecular biological techniques in MABC identification and its pivotal role in guiding therapeutic strategies.

The purpose of this study was to evaluate the application value of targeted next-generation sequencing(tNGS)for rapid detection of Mycobacterium tuberculosis.From September 2021 to October 2024,1 178 clinical samples from hospitalized pa-tients at the Non-tuberculous Mycobacteria Disease Diagnosis and Treatment Center of Guangzhou Chest Hospital were collected,in-cluding 272 sputum samples,871 broncho alveolar lavage fluid(BALF)samples,and 35 other samples.These samples were analyzed with tNGS and the liquid culture+DNA microarray chip method(referred to as the"culture identification method"),and the detection results between methods were compared.The numbers of Mycobacterium tuberculosis complex(MTBC)samples identified with tNGS and the culture identification method were 172 and 127,respectively,and the numbers of non-tuberculous mycobacteria(NTM)samples detected were 517 and 474,respectively.The detection rate of Mycobacterium was 56.88%with tNGS and was significantly lower(49.49%)with the culture identification method(χ2=13.27,P<0.05).For NTM identification,when sputum was used,the detection rate of tNGS was higher than that of the culture identification method(χ2=24.14,P<0.05).However,when BALF was used,no significant difference in detection rates was observed between tNGS and the culture identification method(χ2=3.97,P=0.06).When different sample types from the same patient were analyzed with tNGS for NTM,BALF was found to be a better choice than sputum(χ2=4.05,P<0.05).However,with the culture identification method,we observed no significant difference between sample types(χ2=2.72,P=0.10).Furthermore,both the MTBC and NTM sequences detected with tNGS were significantly higher in the culture-positive group than the culture-negative group.With increasing sequence numbers,the proportion of cases in the culture-positive group progressively rose,whereas an inverse trend was observed in the culture-negative group.With clinical diagno-sis as the reference standard,the sensitivity,specificity,and Kappa value of tNGS and the culture identification method in identifying mycobacterial diseases were 85.41%,91.32%,0.74 and 74.73%,93.15%,0.63,respectively.Compared with the culture identifica-tion method,tNGS for rapid detection of Mycobacterium exhibited excellent sensitivity,specificity and consistency,and its timeliness should help clinicians make precise individualized treatment plans earlier.

In recent year，the iatrogenic infections and outbreaks caused by Mycobacterium abscessus complex（MABC）have been increasingly reported. Studies indicate that MABC accounts for 65%-80% of rapidly growing non-tuberculosis Mycobacterium（NTM）-related pulmonary diseases. As a clinically significant NTM pathogen，MABC exhibits rapid proliferation and intrinsic resistance to multiple antimicrobial agents. Notably，the 16S rDNA sequences among MABC subspecies share 100% homology，posing challenges for subspecies-level differentiation. Rapid and precise identification of MABC at the species or subspecies level is therefore critical for effective clinical management and outbreak control. Conventional biochemical methods for NTM identification are time-consuming and prone to inaccuracies，whereas molecular biology techniques offer superior speed and specificity，and have been widely used clinically. This review highlights the clinical implications of molecular biological techniques in MABC identification and its pivotal role in guiding therapeutic strategies.

Objective This study aims to analyze the proteomic characteristics of peripheral blood in patients with lung cancer coexistent with pulmonary tuberculosis(LC-PTB),identify the differential proteins compared with lung cancer(LC)patients,and conduct functional analysis on these proteins.Methods The study included 8 LC-PTB patients and 10 LC patients.The LC patients were newly diagnosed and confirmed by pathology and did not receive any anti-tumor treatment before,while the PTB patients were Mycobacterium tuberculosis positive at the time of sampling.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was applied to perform proteomic mass spectrometry to assess the differential proteins,and then functional analysis was conducted via bioinformatics.Results A total of 5,185 proteins were detected between two groups.Through differential expression screening,190 proteins(58 upregulated and 132 downregulated)were identified to be differentially expressed.Subcellular localization analysis revealed that the differential proteins were mainly concentrated in the cytoplasm,nucleus,and extracellular matrix.KEGG pathway and GO analysis showed the roles of differential proteins in biological processes including immune response,metabolism,and secretion regulation.Protein interaction network analysis highlighted the importance of SORT1,SAR1B,RPS6KB1,VWF,SHC1,SRPRB,CTSD,TARDBP,RPLP0,PSMA2,RPS6,XPO1,PRKACB,and HLA-DRB1 in LC-PTB.Additionally,the expression changes in proteins like ADA2,MAP3K1,and GLS2 might be closely associated with the development of LC-PTB.Conclusions The proteomic profile comprehensively described the proteomic characteristics of LC-PTB and identified numerous differentially expressed proteins,which could provide further clues for research on biological mechanism of LC-PTB.

Objective This study aims to assess the clinical value of sputum and bronchoalveolar lavage fluid examination combined with acid-fast bacilli detection to provide a reference for the diagnosis and treatment of pulmonary tuberculosis.Methods We collected and analyzed relevant test data from patients who underwent smear and/or isolation of sputum and bronchoalveolar lavage fluid for acid-fast bacilli or Mycobacterium detection within the same week from January 2021 to July 2021.The test results'similarities and differences were analyzed.Results Of the 272 patients,the positive rates of sputum smear,alveolar lavage fluid smear,sputum isolation,alveolar lavage fluid isolation(hereinafter referred to as"A""B""C"and"D")were 14.71％(40/272),19.49％(53/272),25.00％(67/268)and 31.90％(74/232),respectively.The positive rate of the four tests as parallel tests was 37.50％(102/272).The result modes of A+C+,A-C+,A+C-,A-C-and A-CN(the"+""-"and"N"in the super-script stood for"positive""negative"and"undetected")accounted for 14.71％(40/272),13.97％(38/272),0,69.85％(190/272),1.47％(4/272)respectively,and the result modes of B+D+,B-D+,B+D-,B-D-and B-DN accounted for 19.12％(52/272),8.82％(24/272),0.37％(1/272),56.99％(155/272),14.71％(40/272).The percentages of these re-sult modes of A+B+,A+B-,A-B+and A-B-were 14.71％(40/272),0,4.78％(13/272),80.51％(219/272),respec-tively.The percentages of these result modes of A+D+,A+D-,A+DN,A-D+,A-D-,A-DN,AND+,AND-and ANDN were 19.12％(52/272),5.51％(15/272),4.04(11/272),8.09％(22/272),51.74％(140/272),10.29％(28/272),0.74％(2/272),0.37％(1/272),and 0.37％(1/272),respectively.Conclusion Compared with more common sputum tes-ting,for acid-fast bacteria,performing bronchoalveolar lavage fluid testing for acid-fast bacteria in alveolar lavage fluid can signifi-cantly improve etiological diagnostic performance for tuberculosis,which is worth promoting extensively in clinical practice.

Mycobacterium abscessus complex (MABC), a rapidly growing nontuberculous mycobacterium, has received increasing attention worldwide due to its rising isolation rate. The similarity of symptoms between MABC pulmonary disease and tuberculosis, different treatment methods required by different subtypes, as well as high levels of innate, adaptive and acquired antibiotic resistance, make MABC treatment more difficult and lead to unfavorable clinical outcomes of patients. This article reviews the basic characteristics, common antibiotic resistance mechanisms, as well as diagnosis and treatment of MABC, to provide reference for future research and clinical treatment of MABC lung disease.