ABSTRACTIntroductionThe objectives of the study are to understand the drug‐resistant situation and trend of tuberculosis patients in Yuexiu District, Guangzhou City, from 2013 to 2022, and to provide a scientific basis for the development of rational drug‐resistant tuberculosis prevention and control strategies in Guangzhou City.MethodsAll patients who were diagnosed with active tuberculosis in Guangzhou Chest Hospital from January 1, 2013 to December 31, 2022 were collected as study subjects, and a total of 5191 patients were enrolled in the study. Comprehensive data on the basic characteristics, diagnostic, and therapeutic information of the study subjects were collected. Sputum specimens were subjected to smear, isolation, and culture. Culture‐positive strains of bacteria were identified by bacterial groups. A total of 1659 strains of Mycobacterium tuberculosis (MTB) isolates were obtained. The drug susceptibility test was carried out using the proportionality method on the MTB isolates for nine types of antituberculosis medicines: isoniazid (INH), rifampicin (RFP), ethambutol (EMB), streptomycin (Sm), kanamycin (Km), ofloxacin (Ofx), capreomycin (Cm), propylthioisonicotinamide (Pto), and p‐aminosalicylic acid (PAS). A comparative analysis of the resistance patterns among the strains was conducted.ResultsA total of 1659 patients with MTB were cultured, revealing 438 drug‐resistant cases. Among these, 255 were monoresistant, 121 were polyresistant, and 62 were multidrug resistant. The overall resistance rate was 26.40% (438/1659), with mono‐resistance rate at 15.37% (255/1659), polyresistance rate at 7.29% (121/1659), and multidrug resistance rate at 3.74% (62/1659). In descending order, the resistance rates of MTB isolates to any of the nine antituberculosis drugs were Sm (12.24%, 203/1659), INH (9.22%, 153/1659), EMB (7.35%, 122/1659), RFP (6.99%, 116/1659), PAS (3.25%, 54/1659), Pto (3.13%, 52/1659), Ofx (2.71%, 45/1659), Cm (2.17%, 36/1659), and Km (2.17%, 36/1659). The differences in resistance rates were statistically significant (p < 0.01), with Sm exhibiting the highest resistance rate and Km the lowest.In the primary treatment group, 388 patients (25.55%) were drug resistant, while 50 patients (35.46%) in the retreatment group were drug resistant. Thirty‐nine patients (2.57%) in the primary treatment group were multidrug resistant, compared to 23 patients (16.31%) in the retreatment group. The resistance rate and multidrug resistance rate of isolates from retreatment patients were significantly higher than primary treatment patients (p < 0.05).ConclusionsThe problem of drug‐resistant tuberculosis transmission in Guangzhou requires attention, and drug‐resistant screening should be further increased to effectively control the source of infection.

Treatment of Mycobacterium abscessus (Mab) infection is a major challenge due to its intrinsic resistance to most available drugs. It is thus imperative to find new anti-Mab drugs. In this study, we investigated the activity and intrinsic resistance mechanism of echinomycin (ECH) against Mab. ECH is active against Mab (MIC: 2 µg/mL). The embC gene knockout strain (Mab ΔembC ) showed hyper-sensitive to ECH (MIC: 0.0078-0.0156 µg/mL). The MICs of ECH-resistant strains screened based on the Mab ΔembC strain were 0.25-1 µg/mL. Mutations were found in the EmbB, including Asp306Ala, Asp306Asn, Arg350Gly, Val555Ile, and Gly581Ser, which led to increased resistance to ECH when overexpressed in Mab ΔembC individually (0.25-0.5 µg/mL). The EmbB mutants edited by the CRISPR/Cpf1 system became more resistant to ECH (MIC: 0.25-0.5 µg/mL). The permeability of gene-edited and overexpressed Mab strains was reduced, as shown by the ethidium bromide accumulation assay, but it was still significantly higher than that of the parent Mab. To summarize, our study demonstrates that ECH has a strong anti-Mab activity and confirms that EmbB and EmbC are related to the sensitivity of Mab to ECH. EmbB mutation may partially compensate for the function of EmbC. Impact Statement Mycobacterium abscessus (Mab) is a rapidly growing, intrinsic multidrug-resistant Mycobacterium. This study demonstrated that echinomycin (ECH) has potent antibacterial activity against Mab, and the mechanism of ECH resistance to Mab is related to EmbB and EmbC. EmbB and EmbC can alter the sensitivity of Mab to ECH by altering the permeability of its cell wall. In addition, there is a functional complementary evolution between EmbB and EmbC to regulate sensitivity to ECH. Overall, our study provides a novel anti-Mab drug candidate ECH and a scientific foundation for developing effective strategies to prevent and control Mab.

Treatment of Mycobacterium abscessus (Mab) infections is very challenging due to its intrinsic resistance to most available drugs. Therefore, it is crucial to discover novel anti-Mab drugs. In this study, we explored an intrinsic resistance mechanism through which Mab resists echinomycin (ECH). ECH showed activity against Mab at a minimum inhibitory concentration (MIC) of 2 µg/ml. A ΔembC strain in which the embC gene was knocked out showed hypersensitivity to ECH (MIC: 0.0078–0.0156 µg/ml). The MICs of ECH-resistant strains screened with reference to ΔembC ranged from 0.25 to 1 µg/ml. Mutations in EmbB, including D306A, D306N, R350G, V555I, and G581S, increased the Mab’s resistance to ECH when overexpressed in ΔembC individually (MIC: 0.25–0.5 µg/ml). These EmbB mutants, edited using the CRISPR/Cpf1 system, showed heightened resistance to ECH (MIC: 0.25–0.5 µg/ml). The permeability of these Mab strains with edited genes and overexpression was reduced, as evidenced by an ethidium bromide accumulation assay, but it remained significantly higher than that of the parent Mab. In summary, our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab’s sensitivity to ECH. Mutation in EmbB may partially compensate for a loss of EmbC function.

Background: Tuberculosis (TB) remains a significant global health emergency caused by Mycobacterium tuberculosis (Mtb). The epidemiology, transmission, genotypes, mutational patterns, and clinical consequences of TB have been extensively studied worldwide, however, there is a lack of information regarding the epidemiology and mutational patterns of Mtb in Pakistan, specifically concerning the prevalence of multi-drug resistant TB (MDR-TB). Methods: This study aimed to investigate the incidence of Mtb and associated mutational patterns using the line probe assay (LPA). Previous studies have reported a high frequency of mutations in the rpoB, inhA, and katG genes, which are associated with resistance to rifampicin (RIF) and isoniazid (INH). Therefore, the current study utilized LPA to detect mutations in the rpoB, katG, and inhA genes to identify multi-drug resistant Mtb. Results: LPA analysis of a large pool of Mtb isolates, including samples from 241 sputum-positive patients, revealed that 34.85% of isolates were identified as MDR-TB, consistent with reports from various regions worldwide. The most prevalent mutations observed were rpoB S531L and inhA promoter C15T, which were associated with resistance to RIF and INH, respectively. Conclusions: This study highlights the effectiveness of GenoType MTBDRplus and MTBDRsl assays as valuable tools for TB management. These assays enable rapid detection of resistance to RIF, INH, and fluoroquinolones (FQs) in Mtb clinical isolates, surpassing the limitations of solid and liquid media-based methods. The findings contribute to our understanding of MDR-TB epidemiology and provide insights into the genetic profiles of Mtb in Pakistan, which are essential for effective TB control strategies.

ABSTRACT The polymorphism at amino acid 17 of quinolone resistance-determining region of GyrA has been stated with a potential role in fluoroquinolone susceptibility in different mycobacterial species. However, no study has provided dependable evidence so far. Here, we verified that gene-edited Mycobacterium abscessus mutants bearing Ser/Gly at this position were more susceptible to fluoroquinolones than their parent strain and the revertant that supports mycobacteria containing Ser/Gly at this position were more susceptible to fluoroquinolones than those containing Ala. IMPORTANCE Fluoroquinolones (FQs) play a key role in the treatment regimens against tuberculosis and non-tuberculous mycobacterial infections. However, there are significant differences in the sensitivities of different mycobacteria to FQs. In this study, we proved that this is associated with the polymorphism at amino acid 17 of quinolone resistance-determining region of Gyrase A by gene editing. This is the first study using CRISPR-associated recombination for gene editing in Mycobacterium abscessus to underscore the contribution of the amino acid substitutions in GyrA to FQ susceptibilities in mycobacteria.

现代临床诊疗决策对医学检验的依赖日益增加,但目前超过40%的结核病例未获得病原学确诊证据,且普遍存在诊疗延误.准确、快速的实验技术与方法一直为人们所期盼,本文拟从潜伏感染、临床诊断、精准治疗和流行控制等方面探讨结核病临床诊疗和流行控制实践中对医学检验的现实需求与存在问题,同时也就如何走出困境简要给出一些建议.

Introduction Infections caused by non-tuberculosis mycobacteria are significantly worsening across the globe. M. fortuitum complex is a rapidly growing pathogenic species that is of clinical relevance to both humans and animals. This pathogen has the potential to create adverse effects on human healthcare. Methods The MF GZ001 clinical strain was collected from the sputum of a 45-year-old male patient with a pulmonary infection. The morphological studies, comparative genomic analysis, and drug resistance profiles along with variants detection were performed in this study. In addition, comparative analysis of virulence genes led us to understand the pathogenicity of this organism. Results Bacterial growth kinetics and morphology confirmed that MF GZ001 is a rapidly growing species with a rough morphotype. The MF GZ001 contains 6413573 bp genome size with 66.18 % high G+C content. MF GZ001 possesses a larger genome than other related mycobacteria and included 6156 protein-coding genes. Molecular phylogenetic tree, collinearity, and comparative genomic analysis suggested that MF GZ001 is a novel member of the M. fortuitum complex. We carried out the drug resistance profile analysis and found single nucleotide polymorphism (SNP) mutations in key drug resistance genes such as rpoB, katG, AAC(2')-Ib, gyrA, gyrB, embB, pncA, blaF, thyA, embC, embR, and iniA. In addition, the MF GZ001strain contains mutations in iniA, iniC, pncA, and ribD which conferred resistance to isoniazid, ethambutol, pyrazinamide, and para-aminosalicylic acid respectively, which are not frequently observed in rapidly growing mycobacteria. A wide variety of predicted putative potential virulence genes were found in MF GZ001, most of which are shared with well-recognized mycobacterial species with high pathogenic profiles such as M. tuberculosis and M. abscessus. Discussion Our identified novel features of a pathogenic member of the M. fortuitum complex will provide the foundation for further investigation of mycobacterial pathogenicity and effective treatment.

Background Current clinical tests for mycobacterial pulmonary diseases (MPD), such as pulmonary tuberculosis (PTB) and non-tuberculous mycobacteria pulmonary diseases (NTM-PD), are inaccurate, time-consuming, sputum-dependent, and/or costly. We aimed to develop a simple, rapid and accurate breath test for screening and differential diagnosis of MPD patients in clinical settings.Methods Exhaled breath samples were collected from 93 PTB, 68 NTM-PD and 4 PTB&NTM-PD patients, 93 patients with other pulmonary diseases (OPD) and 181 healthy controls (HC), and tested using the online high-pressure photon ionisation time-of-flight mass spectrometer (HPPI-TOF-MS). Machine learning models were trained and blindly tested for the detection of MPD, PTB, NTM-PD, and the discrimination between PTB and NTM-PD, respectively. Diagnostic performance was evaluated by metrics of sensitivity, specificity, accuracy, and area under the receiver operating characteristic curve (AUC).Results The breath PTB detection model achieved a sensitivity of 73.5%, a specificity of 85.8%, an accuracy of 82.9%, and an AUC of 0.895 in the blinded test set (n=141). The corresponding metrics for the NTM-PD detection model were 86.4%, 93.2%, 92.1% and 0.972, respectively. For distinguishing PTB from NTM-PD, the model also achieved good performance with sensitivity, specificity, accuracy, and AUC of 85.3%, 81.8%, 83.9% and 0.947, respectively. 22 potential breath biomarkers associated with MPD were putatively identified and discussed, which included 2-furanmethanol, ethanol, 2-butanone, etc.Conclusions The developed breathomics-based MPD detection method was demonstrated for the first time with good performance for potential screening and diagnosis of PTB and NTM-PD using a refined operating procedure on the HPPI-TOF-MS platform.### Competing Interest StatementThe authors have declared no competing interest.### Funding StatementThis work was supported by Science and Technology Program of Guangzhou (2023A03J0991, 2023A03J0539), Guangzhou High Level Clinical Key Specialty - Tuberculosis, and Guangzhou Medical Key Disciplines - Tuberculosis (2021-2023).### Author DeclarationsI confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.YesThe details of the IRB/oversight body that provided approval or exemption for the research described are given below:Ethics Review Approval of Medical Ethics Committee of Guangzhou Chest Hospital (No. 2022-65): The ethics committee has reviewed the study protocol and found that it is in line with the relevant laws and regulations and the ethical requirements of medical research, and agreed to carry out the study. Patients should be properly informed and sign informed consent forms during project implementation, and patient information should be kept confidential. The Ethics Committee should be notified in a timely manner of any suspension/early termination of the project, and any changes to the implementation plan etc. during the study must be notified to the Ethics Committee in a timely manner for record or re-examined for approval before proceeding, and the Ethics Committee's final report or final summary must be submitted in a timely manner for record.I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.YesI understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as [ClinicalTrials.gov][1]. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).YesI have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.YesAll data produced in the present study are available upon reasonable request to the authors. [1]: https://ClinicalTrials.gov

目的:了解广州市中学和大学新生结核分枝杆菌潜伏感染(latent tuberculosis infection,LTBI)的情况,为广州市学校结核病防控工作和策略制定提供参考依据.方法:选取广州市中学和大学共17所学校作为试点单位,对试点单位2018-2021年共计17 632名入学新生(包括初高中、专科、高等院校的一年级学生)根据知情同意的原则进行结核菌素皮肤试验(tuberculin skin test,TST),对15岁以上学生同时加做胸部X线摄片检查;TST中度及以上阳性者(硬结平均直径≥10 mm)再进一步加做γ-干扰素释放试验(interferon gamma release assay,IGRA);TST和IGRA均为阳性或胸部X线摄片发现肺部异常阴影者,再进行痰涂片、痰培养等结核病辅助诊断检查.使用描述性流行病学方法对筛查结果进行分析.结果:实际筛查学生17 204名,筛查率为97.57％(17 204/17 632).TST 阳性者共 4449 名,阳性率为 25.87％(4449/17 199),一般阳性率为 15.98％(2748/17 199),中度阳性率为 7.65％(1315/17 199),强阳性率为 2.24％(386/17 199).高中组 TST 阳性率(33.81％,1408/4165)最高,初中组TST阳性率(30.58％,329/1076)次之,大学组TST阳性率(22.68％,2712/11 958)最低,组间差异有统计学意义(x2=353.897,P＜0.001).男生TST阳性率为26.68％(2789/10 452),明显高于女生的24.60％(1660/6747),差异有统计学意义(x2=35.855,P＜0.001).1701名TST中度及以上阳性者(即硬结平均直径≥10 mm)加做IGRA检测,IGRA阳性者244例,TST中度及以上阳性+IGRA阳性率为1.42％(244/17 199).筛查发现7例学生胸片异常,其中1例TST阴性(IGRA未做),6例TST阳性(其中5例IGRA阳性,1例IGRA阴性).最终确诊肺结核5例,肺结核检出率为29.06/10万(5/17 204).结论:广州市学生结核感染情况不容忽视,应加强初中、高中新生入学结核病体检工作,及时发现学生结核病患者和LTBI者.TST和IGRA联合筛查有助于提高筛查的准确性和防控效率,符合成本效益原则.

目的:分析广州市耐药肺结核高危人群耐药情况及特征,以及耐药发生的影响因素.方法:从"中国疾病预防控制信息系统"的子系统"结核病管理信息系统"中搜集广州市2014年1月1日至2019年12月31日期间登记的耐药肺结核高危人群资料,包括社会人口学特征、耐药筛查结果等资料,最终纳入2155例研究对象的相关信息.分析研究对象对5种抗结核药品[异烟肼(isoniazid,INH)、利福平(rifampicin,RFP)、乙胺丁醇(ethambu-tol,EMB)、氧氟沙星(ofloxacin,Ofx)、卡那霉素(kanamycin,Km)]的耐药情况、耐药顺位、耐药谱,以及影响耐药发生的因素.结果:2155例研究对象中有768例耐药,总耐药率为35.64％,单耐药率、耐多药率、广泛耐药率分别为10.39％(224/2155)、24.36％(525/2155)、0.88％(19/2155).研究对象对5种抗结核药品的耐药顺位由高到低依次为 INH(31.97％,689/2155)、RFP(29.05％,626/2155)、EMB(10.72％,231/2155)、Ofx(7.75％,167/2155)、Km(2.55％,55/2155).耐 1 种药品至耐 5 种药品的比例分别为 24.22％(186/768)、39.97％(307/768)、24.87％(191/768)、9.38％(72/768)和 1.56％(12/768).耐 1 种药品者中,以耐 INH 最多,占 15.23％(117/768);耐 2 种药品者中,以耐INH+RFP最多,占35.94％(276/768);耐3种药品者中,以耐INH+RFP+EMB最多,占16.80％(129/768);耐 4 种药品者中,以耐 INH+RFP+EMB+Ofx最多,占 6.51％(50/768).多因素 logistic 回归分析结果显示,≥65岁组患者耐药发生风险是＜25岁组的37.9％(OR=0.379,95％CI:0.226～0.634);职业分类中,商业服务,教师、医务人员及干部职员,农民,以及其他者耐药发生风险分别是离退休人员的2.419倍(95％CI:1.429～4.096)、2.541 倍(95％CI:1.325～4.873)、1.479 倍(95％CI:1.028～2.127)、6.452 倍(95％CI:4.624～9.003);患者分类中,初治失败、复治失败/慢性患者、复发、其他者耐药发生风险分别是初治2、3个月末痰涂片阳性者的 9.443 倍(95％CI:6.009～14.621)、7.504 倍(95％CI:4.634～12.151)、2.567 倍(95％CI:1.968～3.348)和3.091倍(1.969～4.854).结论:近年来广州市耐药肺结核高危人群中超过1/3出现耐药,耐药形势不容忽视.耐药肺结核高危人群中要重点关注中青年,关注商业服务、教师、医生、企事业单位职员、农民等从业者,关注初治失败、复治失败/慢性患者、复发等人群.

Background: Tuberculosis (TB) continues to be a major cause of morbidity and mortality worldwide. How-ever, the molecular mechanism underlying immune response to human infection with Mycobacterium tuberculosis (Mtb) remains unclear. Assessing changes in transcript abundance in blood between health and disease on a genome-wide scale affords a comprehensive view of the impact of Mtb infection on the host defense and a reliable way to identify novel TB biomarkers.Methods: We combined expression profiling by array and single cell RNA-sequencing (scRNA-seq) via 10X Genomics platform to better illustrate the immuno-related transcriptional signature of TB and explore potential diagnostic markers for differentiating TB from latent tuberculosis infection (LTBI) and healthy control (HC). Findings: Pathway analysis based on differential expressed genes (DEGs) revealed that immune transcrip-tional profiling could effectively differ TB with LTBI and HC. Following WGCNA and PPI network analysis based on DEGs, we screened out three key immuno-related hub genes (ADM, IFIT3 and SERPING1) highly associated with TB. Further validation found only ADM expression significantly increased in TB patients in both adult and children's datasets. By comparing the scRNA-seq datasets from TB, LTBI and HC, we observed a remarkable elevated expression level and proportion of ADM in TB Myeloid cells, further sup-porting that ADM expression changes could distinguish patients with TB from LTBI and HC. Besides, the hsa-miR-24-3p-NEAT1-ADM-CEBPB regulation pathway might be one of the critical networks regulating the pathogenesis of TB. Although further investigation in a larger cohort is warranted, we provide useful and novel insight to explore the potential candidate genes for TB diagnosis and intervention. Interpretation: We propose that the expression of ADM in peripheral blood could be used as a novel biomarker for differentiating TB with LTBI and HC.(c) 2022 The Authors. Published by Elsevier Ltd on behalf of The British Infection Association. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/ )

The centromere is a vital chromosomal structure that provides all living cells with the ability to faithfully partition their genetic material during mitotic and meiotic cell divisions. It functions by holding newly replicated sister chromatids together, allowing the attachment of spindle microtubules, and orchestrating the ordered movement of chromosomes to the daughter cells. The centromere has also been recognized as a "marshalling station" for a host of "passenger proteins" that appear transiently on the centromere during specific stages of the cell cycle (Earnshaw and Mackay 1994).

Klebsiella pneumoniae is an opportunistic pathogen that is responsible for community acquired infections and nosocomial infections. Antibiotic-resistant K. pneumoniae and/or hypervirulent K. pneumoniae are emerging as a serious threat to public health. For the sake of alleviating and conquering current dilemma, discovery of effective new drugs against K. pneumoniae is a tough challenge. However, traditional anti-K. pneumoniae drug discovery methods cost considerable amount of time, animals, labor and so on. So an efficient technique for in vitro and in vivo drug screening with the least time duration, animals and labor cost is highly needed for the discovery of new effective compounds. Hence, in this study we constructed a selectable marker-free autoluminescent K. pneumoniae (SfAlKp) harboring luxCDABE by combining Tn7 transposon and Xer-dif system. SfAlKp can be used for discovery of new drugs via detecting luminescence intensity as a surrogate marker. The energy-consuming autoluminescent reaction catalyzed by the LuxAB enzymes which use the substrates produced by LuxCDE using the metabolites of the bacteria. Tn7 can insert exogenous genes into the bacterial genome and the DNA fragment in between dif sequences can be recognized and removed by endogenous XerCD recombinases of K. pneumoniae. The drug susceptibility and growth rate of SfAlKp are identical to its parent strain, meanwhile the luminescence intensity and stability are also significant characteristics of SfAlKp. Compared to conventional techniques, the autoluminescence-based measurement is more applicable to high throughput screening for compounds both in vitro as well as in vivo in animal model.

Drug-resistant tuberculosis (DR-TB) poses a new threat to global health; to improve the treatment outcome, therapeutic vaccines are considered the best chemotherapy adjuvants. Unfortunately, there is no therapeutic vaccine approved against DR-TB. Our study assessed the therapeutic efficacy of a recombinant drug-resistant BCG (RdrBCG) vaccine in DR-TB. We constructed the RdrBCG overexpressing Ag85B and Rv2628 by selecting drug-resistant BCG strains and transformed them with plasmid pEBCG or pIBCG to create RdrBCG-E and RdrBCGI respectively. Following successful stability testing, we tested the vaccine's safety in severe combined immune deficient (SCID) mice that lack both T and B lymphocytes plus immunoglobulins. Finally, we evaluated the RdrBCG's therapeutic efficacy in BALB/c mice infected with rifampin-resistant M. tuberculosis and treated with a second-line anti-TB regimen. We obtained M. bovis strains which were resistant to several second-line drugs and M. tuberculosis resistant to rifampin. Notably, the exogenously inserted genes were lost in RdrBCG-E but remained stable in the RdrBCG-I both in vitro and in vivo. When administered adjunct to a second-line anti-TB regimen in a murine model of DR-TB, the RdrBCG-I lowered lung M. tuberculosis burden by 1 log10. Furthermore, vaccination with RdrBCG-I adjunct to chemotherapy minimized lung tissue pathology in mice. Most importantly, the RdrBCG-I showed almost the same virulence as its parent BCG Tice strain in SCID mice. Our findings suggested that the RdrBCG-I was stable, safe and effective as a therapeutic vaccine. Hence, the "recombinant" plus "drugresistant" BCG strategy could be a useful concept for developing therapeutic vaccines against DR-TB.

BACKGROUND:We performed a prospective multicentre diagnostic study to evaluate the combined interferon-γ (IFN-γ) and interleukin-2 (IL-2) release assay for detect active pulmonary tuberculosis (TB) in China.METHODS:Adult patients presenting symptoms suggestive of pulmonary TB were consecutively enrolled in three TB-specialized hospitals. Sputum specimens and blood sample and were collected from each participant at enrolment. The levels of Mycobacterium tuberculosis (MTB)-specific antigen-stimulated IFN-γ and IL-2 were determined using enzyme-linked immunosorbent assay (ELISA).RESULTS:Between July 2017 and December 2018, a total of 3245 patients with symptoms suggestive of pulmonary TB were included in final analysis. Of 3245 patients, 2536 were diagnosed as active TB, consisting of 1092 definite TB and 1444 clinically diagnosed TB. The overall sensitivity and specificity of IFN-γ were 83.8% and 81.5%, respectively. In addition, compared with IFN-γ, the specificity of IL-2 increased to 94.3%, while the sensitivity decreased to 72.6%. In addition, the highest sensitivity was achieved with parallel combination of IFN-γ/IL-2, with a sensitivity of 87.9%, and its overall specificity was 79.8%. The sensitivity of series combination test was 68.5%. Notably, the sensitivity of series combination test in definite TB (72.1%) was significantly higher than that in clinically diagnosed TB (65.8%).CONCLUSION:In conclusion, we develop a new immunological method that can differentiate between active TB and other pulmonary diseases. Our data demonstrates that the various IFN-γ/IL-2 combinations provides promising alternatives for diagnosing active TB cases in different settings. Additionally, the diagnostic accuracy of series combination correlates with severity of disease in our cohort.

继全球“现代结核病控制策略”(1991 2005年)和“遏制结核病策略”(2006-2015年)之后,WHO又于2014年提出了“终止结核病策略”(2016-2035年)[1].其主要目标是:(1)相比2015年,结核病发病率下降90％(＜10/10万)、死亡率下降95％;(2)没有因结核病而面临灾难性支出的家庭.但从2016年以来的实践结果看,人类要实现2035年终止结核病目标极具挑战[2-3].我国作为30个结核病高负担、30个耐药结核病高负担和14个TB/HIV高负担国家之一[2],加上仍是发展中人口大国的现实国情,更是任重而道远.但是,我国正在进行且取得重大战果的抗击新型冠状病毒肺炎的壮阔实践则向世界充分彰显了中国优势、中国速度、中国力量、中国精神,我们认为:只要人们能够直面下述一些重大问题,并着力探索符合中国国情的问题解决方案,在中国实现“终止结核病”之梦并非难以企及.

Tuberculosis (TB) remains a serious global public health problem in the present. TB also affects other sites (extrapulmonary tuberculosis, EPTB), and accounts for a significant proportion of tuberculosis cases worldwide. In order to comprehensively understand epidemiology of EBTB in China, and improve early diagnosis and treatment, we conducted a large-scale multi-center observational study to assess the demographic data and the prevalence of common EPTB inpatients, and further evaluate the prevalence of EPTB concurrent with Pulmonary tuberculosis (PTB) and the associations between multiple EPTB types and gender-age group in China. All consecutive age≥15yr inpatients with a confirmed diagnosis of EPTB during the period from January 2011 to December 2017 were included in the study. The descriptive statistical analysis included median and quartile measurements for continuous variables, and frequencies and proportions with 95% confidence intervals (CIs) for categorical variables. Multinomial logistic regression analysis was used to compare the association of multiple EPTB types between age group and gender. The results showed that the proportion of 15-24 years and 25-34 years in EPTB inpatients were the most and the ratio of male: female was 1.51. Approximately 70% of EPTB inpatients were concurrent with PTB or other types of EPTB. The most common of EPTB was tuberculous pleurisy (50.15%), followed by bronchial tuberculosis (14.96%), tuberculous lymphadenitis of the neck (7.24%), tuberculous meningitis (7.23%), etc. It was found that many EPTB inpatients concurrent with PTB. The highest prevalence of EPTB concurrent with PTB was pharyngeal/laryngeal tuberculosis (91.31%), followed by bronchial tuberculosis (89.52%), tuberculosis of hilar lymph nodes (79.52%), tuberculosis of mediastinal lymph nodes (79.13%), intestinal tuberculosis (72.04%), tuberculous pleurisy (65.31%) and tuberculous meningitis (62.64%), etc. The results from EPTB concurrent with PTB suggested that females EPTB inpatients were less likely to be at higher risk of concurrent PTB (aOR = 0.819, 95%CI:0.803-0.835) after adjusted by age. As age increasing, the trend risk of concurrent PTB decreased (aOR = 0.994, 95%CI: 0.989-0.999) after adjusted by gender. Our study demonstrated that the common EPTB were tuberculous pleurisy, bronchial tuberculosis, tuberculous lymphadenitis of the neck, tuberculous meningitis, etc. A majority of patients with pharyngeal/laryngeal tuberculosis, bronchial tuberculosis, tuberculosis of hilar/mediastinal lymph nodes, intestinal tuberculosis, tuberculous pleurisy, tuberculous meningitis, etc. were concurrent with PTB. Female EPTB inpatients were less likely to be at higher risk of concurrent PTB, and as age increasing, the trend risk of concurrent PTB decreased. The clinicians should be alert to the presence of concurrent tuberculosis in EPTB, and all suspected cases of EPTB should be assessed for concomitant PTB to determine whether the case is infectious and to help for early diagnosis and treatment.

目的探讨住院肺结核患者并发肺外结核的发生情况及其与性别、年龄的关系。方法采用观察性研究方法,由参加过统一培训的调查员从医院信息管理系统（HIS系统）收集2011年1月1日至2017年12月31日我国15省21家医疗机构360 187例住院肺结核患者的性别、年龄,以及结核病灶累及部位等信息,比较分析肺结核患者并发肺外结核的发生情况及其与性别、年龄的关系。结果 360 187例肺结核患者中,男238 910例（66.33%）,女121 277（33.67%）,年龄中位数（四分位数）[M（Q 1 ,Q 3 ）]为47（28,62）岁;42 987例（11.93%）并发肺外结核,并发率依次为结核性脑膜炎[2.72%（9809例）]、颈部淋巴结结核[1.93%（6966例）]、结核性腹膜炎[1.59%（5733例）]、结核性心包炎[0.94%（3399例）]、肠结核[0.94%（3380例）]等。男性肺结核患者并发结核性脑膜炎、颈部淋巴结结核、结核性腹膜炎、结核性心包炎、结核性多浆膜炎、腰椎结核、胸椎结核、胸壁结核的并发率分别为2.44%（5829例）、1.44%（3429例）、1.41%（3376例）、0.90%（2138例）、0.75%（1791例）、0.67%（1604例）、0.64%（1522例）、0.60%（1438例）,均明显低于女性[分别为3.28%（3980例）、2.92%（3537例）、1.94%（2357例）、1.04%（1261例）、0.90%（1093例）、0.79%（960例）、0.76%（924例）、0.66%（805例）]（χ~2=215.235,930.541,144.480,18.061,23.272,16.442,18.585,4.976;P值均＜0.05）。不同年龄组（1～岁组至≥65岁组）肺结核患者并发结核性脑膜炎、颈部淋巴结结核、结核性腹膜炎、肠结核、结核性心包炎、结核性多浆膜炎、腰椎结核、胸椎结核、胸壁结核、咽喉结核的并发率差异均有统计学意义（χ~2=3870.549,2939.502,1830.620,673.372,115.428,319.078,52.512,19.308,439.177,136.619;P值均＜0.05）。除胸椎结核的并发率未呈现出随年龄变化的趋势(χ 趋势 ~2=0.814,P=0.367),结核性心包炎呈现出随年龄增长而增高的趋势(χ 趋势 ~2=62.087,P＜0.001)外,其他肺外结核的发生率均呈现出随年龄增长而降低的趋势（P值均＜0.001）。多因素logistic回归分析结果显示,肺结核并发肺外结核患者的风险女性高于男性[(OR（95%CI）=1.325（1.297～1.353）];其他各年龄组风险均高于≥65岁年龄组[1～岁组、15～岁组、25～岁组、35～岁组、45～岁组、55～岁组的OR（95%CI）值分别为:4.995（4.655～5.360）、2. 481 （2.397～2.568）、2.053 （1. 982～2.126）、1.683 （1.619～1.749）、1.276 （1.228～1.326）、1.109（1.067～1.153）];在控制了性别的影响后,肺结核并发肺外结核患者的风险随年龄的增长而降低[OR（95%CI）=0.817（0.812～0.821）]。结论肺结核患者可并发结核性脑膜炎、颈部淋巴结结核、结核性腹膜炎、结核性心包炎和肠结核等多种肺外结核;且并发肺外结核的风险女性高于男性,并呈现出随年龄的增长而降低的趋势。

In clinical practice, PTB patients have concurrent many types of comorbidities such as pneumonia, liver disorder, diabetes mellitus, hematological disorder, and malnutrition. Detecting and treating specific comorbidities and preventing their development are important for PTB patients. However, the prevalence of most comorbid conditions in patients with PTB is not well described. We conducted a large-scale, multicenter, observational study to elucidate and illustrate the prevalence rates of major comorbidities in inpatients at 21 hospitals in China. The 19 specific comorbidities were selected for analysis in this patient cohort, and stratified the inpatient cohort according to age and gender. A total of 355,929 PTB inpatients were included, with a male:female ratio of 1.98 and the proportion of ≥ 65 years PTB inpatients was the most. Approximately 70% of PTB inpatients had at least one defined type of comorbidity. The prevalence of 19 specific comorbidities in inpatients with PTB was analyzed, with pneumonia being the most common comorbidity. The prevalence of most comorbidities was higher in males with PTB except thyroid disorders, mental health disorders, etc. The prevalence of defined most comorbidities in patients with PTB tended to increase with increasing age, although some specific comorbidities tended to increase initially then decrease with increasing age. Our study describes multiple clinically important comorbidities among PTB inpatients, and their prevalence between different gender and age groups. The results will enhance the clinical aptitude of physicians who treat patients with PTB to recognize, diagnose, and treat PTB comorbidities early.

目的 了解2008-2018年广州市报告新发肺结核流行特征及趋势.方法 通过《中国疾病预防控制信息系统》搜集2008-2018年广州市报告新发肺结核患者的性别、年龄、职业、地区分布、病原学分类等资料,共计144 473例.2008-2018年广州市各年度及各区常住人口信息来源于广州市统计年鉴.对2008-2018年广州市报告新发肺结核的流行病学特征进行描述性分析.结果 2008-2018年广州市报告新发肺结核患者共计144 473例,年均报告发病率为100.14/10万,报告发病率由2008年的148.44/10万(16 556/11 153 400)下降至2018年的67.58/10万(10 072/14 904 400),差异有统计学意义(x2趋势=6849.75,P＜0.05).2008-2018年广州市报告新发肺结核患者中,男性年均报告发病率为135.65/10万(100 003/73 722 700),年均递减率为7.56％;女性年均报告发病率为63.04/10万(44 470/70547 300),年均递减率为7.38％,男女性间年均报告发病率差异有统计学意义(x2=18 999.59,P＜0.05);各年龄组中25～岁组报告新发患者最多,占23.88％(34 505/144 473);患者职业以家政、家务及待业人群占比最高(23.82％,34 416/144 473).涂阳肺结核患者报告发病率呈逐年下降趋势,从2008年的41.31/10万(4608/11 153 400)下降至2018年的23.14/10万(3449/14 904 400).结论 广州市报告新发肺结核疫情呈逐年下降趋势,需加强男性,青壮年,家政、家务及待业人员的结核病防控工作.