评价显微镜观察药物敏感性技术(MODS)快速检测肺泡灌洗液中结核分枝杆菌对菌阴肺结核的临床诊断价值

目的:探讨肺泡灌洗液中结核分枝杆菌对菌阴肺结核临床诊断过程中显微镜观察药物敏感性技术(MODS)的应用价值。方法:整理2017年3月~2019年3月本院100份肺泡灌洗液标本,对其分别实施MODS、罗氏培养法检测,回顾分析检测结果。结果:与罗氏培养法相比,MODS对结核分枝杆菌的阳性检出时间明显缩短(P<0.05);MODS对结核分枝杆菌的阳性检出率高于罗氏培养法对应的阳性检出率(P<0.05)。结论:肺泡灌洗液中结核分枝杆菌对菌阴肺结核临床诊断期间通过应用MODS技术,可缩短诊断时间,提高阳性检出,保证菌阴性肺结核的诊断准确率。     

荧光定量PCR和FISH技术诊断肺结核的价值比较

目的比较荧光定量PCR和FISH技术对肺结核诊断的敏感性和特异性。方法共收集300例明确诊断为肺结核的患者作为阳性对照组,以及收集300例健康志愿者(明确排除肺结核患者)做为阴性对照组。收集800例新收临床疑似肺结核患者做为研究组。收集该800例患者的肺活检标本以及同期的痰液标本,其中痰液进行涂片、结核杆菌分离培养以及结核杆菌DNA荧光定量PCR检测和FISH检测。比较荧光定量PCR和FISH技术对肺结核诊断阳性符合率和阴性符合率。结果 1FISH技术检测结核杆菌阳性符合率为99.3%;荧光定量PCR检测结核杆菌阳性符合率为97.3%。两者阳性符合率存在统计学差别(p=0.015)。2FISH技术检测结核杆菌阴性符合率为98.2%,荧光定量PCR技术检测结核杆菌阴性符合率为96.9%。两者阴性符合率存在统计学差别(p=0.007)。结论与荧光定量PCR比较,FISH对肺结核诊断具有较高的阳性符合率和阴性符合率。     

不同样本在结核分歧杆菌检测中的应用观察

目的:观察不同类型样本对结核分枝杆菌检出率的差异。方法:选取2018年1～2019年1月确诊为结核病的患者64例,均获取痰液、肺泡灌洗液、尿液及血液1.25%。其中肺泡灌洗液标本检出率明显高于痰液、尿液、血液标本,差异均有统计学意义(P均0.05);痰液标本检出率明显高于尿液或血液(P均0.05);尿液标本检出率与血液比较差异样本,采用荧光定量PCR法对各标本结核分枝杆菌进行检测。结果:64例肺结核患者痰液、肺泡灌洗液、尿液、血液标本经荧光定量PCR技术检测,结核分枝杆菌检出率分别为70.31%、98.44%、29.69%,无统计学意义(P0.05)。结论:不同样本结核分歧杆菌检测的阳性率存在明显差异,以肺泡灌洗液最高,其次为痰液,尿液及血液最低。在肺结核诊断中,应综合多种途径进行诊断,以避免漏诊。     

多指标联合检测在菌阴肺结核诊断中的价值研究

目的研究痰或肺泡灌洗液荧光定量TB-DNA、结核菌噬菌体检测结核菌及T-SPOT多指标联合检测在菌阴肺结核诊断中价值。方法选取2010年10月~2012年10月在本院诊疗的患者223例,分为初治的菌阴肺结核组108例、初治的菌阳肺结核组55例和非肺结核病组60例。对入选患者进行常规检测,同时进行血T-SPOT、结核菌噬菌体检测结核菌和痰或肺泡的灌洗液TB-DNA;对患者的检测结果记录、分析,研究以上3项检查的单项特异性和敏感性,及3项检测结合的诊断价值。结果 3项检查中每个项目检查的特异性均达到了90%以上,菌阴肺结核组和菌阳肺结核组的阳性人数都远大于非肺结核组的人数,3组比较差异明显;联合检测指标的增加,菌阴肺结核中的阳性检出率也随之增加。有T-SPOT参与的检测其阳性率都高于没有T-SPOT参与的检测,差异有统计学意义(P0.05);联合检查在菌阴肺结核组和非肺结核组同时阳性的检出率差异有统计学意义(P0.05)。结论 T-SPOT的检查、结核菌噬菌体检测结核菌和TB-DNA的检查多个项目指标联合检测比单一的检测更能提高菌阴肺结核的诊断敏感性,其中含有T-SPOT检查参与的结合检查更可以提升菌阴肺结核的检查率。     

三种方法检测涂阴肺结核患者痰液和灌洗液结核分枝杆菌结果分析

目的比较三种不同检测方法对涂阴肺结核患者痰液和支气管肺泡灌洗液结核分枝杆菌检测结果,探讨如何提高涂阴患者的病原学阳性率。方法选取2015年7月至2017年6月,衢州市人民医院住院患者117例,分别采用浓缩涂片抗酸染色、液体培养和Xpert MTB/RIF法检测痰液和肺泡灌洗液结核分枝杆菌,比较同种样本不同检测方法的阳性率。结果 117例涂阴患者痰液样本Xpert MTB/RIF检测、浓缩涂片抗酸染色、分枝杆菌液体培养阳性率分别为23.08%(27/117)、4.27%(5/117)、21.37%(25/117),117例涂阴患者肺泡灌洗液样本Xpert MTB/RIF检测、浓缩涂片抗酸染色、分枝杆菌液体培养阳性率分别为45.30%(53/117)、17.09%(20/117)、44.44%(52/117)。同种样本比较Xpert MTB/RIF和液体培养法阳性率高于浓缩涂片法,差异有统计学意义(χ~2=5.96,P0.05)。不同种类样本比较,肺泡灌洗液3种方法阳性检出率均高于痰液同种方法,差异有统计学意义(χ~2=5.13,P0.05)。结论对涂阴患者的样本进行液体培养和Xpert MTB/RIF检测可提高病原学阳性率。肺泡灌洗液检测能进一步提高阳性检出率。     

痰标本中结核杆菌分泌蛋白抗原检测在活动性肺结核诊断中的应用价值

目的了解痰标本中结核杆菌分泌蛋白抗原检测在活动性肺结核诊断中的应用价值。方法选取109例确诊肺结核患者作为实验组,对照组选取住院肺部疾病患者27例和医院职工查体健康者71名。采用痰厚涂片抗酸染色检测、痰结核杆菌分泌蛋白抗原检测、痰结核杆菌Gene Xpert检测进行实验。数据采用SPSS 17.0进行统计分析。结果TBAg快速检测敏感度和特异度分别为50.5%和86.7%,与Gene Xpert MTB/RIF法检测结果基本一致,阳性率分别为50.5%和55.0%。TBAg检测在Gene Xpert MTB/RIF法阳性组的阳性检出率为61.7%,而在Gene Xpert MTB/RIF法阴性组的阳性检出率为32.7%。TBAg阳性55例,平均用药时间为(32.3±11.4)d;TBAg检测阴性54例,平均用药时间为(28.2±9.6)d。结论结核杆菌分泌蛋白抗原快速检测技术直接检测痰标本,操作简便、快速、敏感度和特异度较高,对早期诊断活动性肺结核具有较大价值。     

肺泡灌洗液与血液T细胞斑点试验对肺结核的诊断价值比较

目的对肺泡灌洗液、血液T细胞斑点试验诊断肺结核的临床价值进行对比研究。方法将2015年6月—2016年9月期间,我院接收的疑似肺结核病但还未接受相关药物治疗的92例患者纳入此次研究当中,全部给予肺泡灌洗液检查和血液T细胞斑点检查,对比两种检查方法的临床诊断价值。结果经临床确诊,疑似患者中有47例被确诊为肺结核患者。45例被确诊为非肺结核患者。肺泡灌洗液检查结果,肺结核诊断的敏感度为85.11%,非肺结核诊断的特异性为97.78%;血液T细胞斑点试验结果,肺结核诊断的敏感度为19.15%,非肺结核诊断的特异性为80.00%;肺泡灌洗液检查的敏感度和特异均显著高于血液T细胞斑点试验,差异有统计学意义(P<0.05)。肺泡灌洗液检查下的阳性预测值、阴性预测值和准确性分别为85.11%、86.27%和91.30%,均显著高于血液T细胞斑点试验检查的50.00%、48.65%、48.91%,差异有统计学意义(P<0.05)。结论与血液T细胞斑点试验相比,采用肺泡灌洗液检查进行肺结核的诊断,敏感度和特异性均更高;诊断的阳性预测值、阴性预测值以及准确性也更高,在结核病的诊断中的应用价值较高,能够作为临床诊断和治疗的有效参考。     

肺泡灌洗液二代基因测序在鹦鹉热衣原体感染肺炎诊断中的应用

目的 探究肺泡灌洗液二代基因测序(NGS)技术在鹦鹉热衣原体感染肺炎诊断中的应用价值。方法 选择2019年3月-2021年4月东莞市人民医院收治的13例鹦鹉热衣原体感染肺炎患者,收集其肺泡灌洗液进行传统病原检测和二代基因测序技术病原检测,同时采集患者的痰样本进行二代基因测序技术检测,分析不同检测方法对鹦鹉热衣原体感染肺炎的诊断效能。结果 肺泡灌洗液NGS对鹦鹉热衣原体的阳性检出率显著高于传统病原检测和痰样本NGS(P<0.05)。肺泡灌洗液NGS、痰样本NGS的检测结果回报耗时显著短于传统病原测定(P<0.05)。结论 肺泡灌洗液NGS技术能够显著提高鹦鹉热衣原体感染肺炎的诊断准确率,且检测结果回报快,可为临床制定合理的抗感染治疗方案提供精准的依据,进而有效改善患者的临床结局,减少其治疗费用。     

痰涂片检查联合结核杆菌培养对肺结核的诊断价值

目的:研究痰涂片检查联合结核杆菌培养对肺结核的诊断价值。方法:选取时间节点为:2017年3月～2018年5月来我院的100例肺结核患者作为研究对象,对照组患者采用痰涂片进行检测诊断,观察组患者采用痰涂片联合结核杆菌培养进行诊断,对两组患者检测后的阳性率进行观察和比较。结果:观察组患者检测后的阳性率优于对照组患者检测的阳性率,P0.05。结论:对肺结核患者采用痰涂片检查联合结核杆菌培养进行诊断,可以提高患者肺结核的阳性检出率,值得被临床推广和应用。     

免疫胶体金法检测支气管肺泡灌洗液MPB64对肺结核的诊断价值

目的探讨免疫胶体金法检测支气管肺泡灌洗液(BALF)结核杆菌特异性分泌抗原MPB64在肺结核诊断中的应用价值。方法对113例肺结核患者和80例非结核患者的BALF进行免疫胶体金法检测MPB64、荧光定量PCR法检测结核分枝杆菌DNA及改良酸性罗氏培养基法结核分枝杆菌培养,比较分析3种方法的检测结果。结果 MPB64免疫胶体金法与荧光定量PCR法、改良酸性罗氏培养基法的符合率分别为58.03%和57.51%;以临床诊断结果为参照,免疫胶体金法检测MPB64的敏感度、特异度与准确度分别为61.95%、87.50%、72.54%。以荧光定量PCR法为参照,免疫胶体金法检测MPB64的敏感度、特异度与准确度分别为49.30%、63.11%、58.03%;以改良酸性罗氏培养基法为参照,免疫胶体金法检测MPB64的敏感度、特异度与准确度分别为48.21%、61.31%、57.51%。结论免疫胶体金法检测MPB64应用于支气管肺泡灌洗液对肺结核的临床诊断有辅助诊断价值,但检测的假阳性、假阴性值得进一步分析研究。     

Mycobacterium tuberculosis arabinomannan-protein conjugates protect against tuberculosis

Lipoarabinomannan (LAM) is a major structural surface component of mycobacteria. Arabinomannan (AM) oligosaccharides derived from LAM of Mycobacterium tuberculosis H37Rv were isolated and covalently conjugated to tetanus toxoid (TT) or to short-term culture filtrate proteins (antigen 85B (Ag85B) or a 75kDa protein) from M. tuberculosis strain Harlingen. The different AM oligosaccharide (AMOs)-protein conjugate vaccine candidates proved to be highly immunogenic, inducing boosterable IgG responses against the AMOs portion of the conjugates in rabbits and guinea-pigs. Proliferation of T-cells from C57BL/6 mice immunized with the conjugates was seen upon in vitro stimulation with PPD. In C57BL/6 mice subcutaneous immunization with the AMOs-antigen 85B conjugate in alum provided significant protection compared to sham (alum only) immunized mice (P < 0.021) as estimated by long term survival against intravenous challenge with 10(5) M. tuberculosis H37Rv. Subcutaneous immunization followed by nasal boost with an AMOs-TT conjugate in Eurocine L3 adjuvant provided high (P < 0.025) protection as determined by long term survival after intranasal challenge with 10(5) virulent M. tuberculosis strain Harlingen. This level of protection was comparable to that obtained with the conventional live attenuated BCG vaccine. In guinea-pigs, immunization with AMOs-Ag85B in Eurocine L3 adjuvant followed by aerogenic challenge with M. tuberculosis H37Rv resulted in increased survival and reduced pathology in lungs and spleens relative to non-immunized animals.     

Immunology of tuberculosis

Various T cells and macrophages as well as cytokines are involved in the immunopathogenesis of tuberculosis (TB). A better understanding of immunology of TB can not only lead to the discovery of new immunodiagnostic tools, accelerate and facilitate the assessment of new therapeutic methods, but also find new treatment regimens. In this highlight topic we cover the latest developments in the role of T cells, macrophages, Natural killer (NK) cells, invariant NK T (iNKT) cells and γδ T cells with TB infection. Histologically, TB displays exudative inflammation, proliferative inflammation and productive inflammation depending on the time course. T cells first recognize antigen within the mycobacterially-infected lung, and then activate, differentiate, but the first T cell activation occurs in the draining lymph nodes of the lung. When protective T cells reach sufficient numbers, they can stop bacterial growth. Except for T cells, neutrophils also participate actively in defense against early-phase TB. NK cells are innate lymphocytes which are a first line of defense against mycobacterial infection. Human NK cells use the NKp46, NCRs and NKG2D receptors to lyse Mycobacterium TB-infected monocytes and alveolar macrophages. NK cells produce not only interferon-γ, but also interleukin (IL)-22, which is induced by IL-15 and DAP-10. iNKT cells show different phenotypes and functions. Many iNKT cells are CD4+, few iNKT cells are CD8+, while an additional fraction of iNKT cells are negative for both CD4 and CD8. γδ T cells represent an early innate defense in antimycobacterial immunity. Studies done in humans and animal models have demonstrated complex patterns of γδ T cell immune responses during chronic TB. Human alveolar macrophages and monocytes can serve as antigen presentation cells for γδ T cells. Furthermore, the predominance of Vγ9Vδ2 T cells in TB has been confirmed.     

Treatment of tuberculosis

Even though children with tuberculosis (TB) tend to be noninfectious and not a risk to the community, they need to be treated. Proper treatment will cure them quickly. To prevent resistance, the anti-TB drugs (isoniazid, rifampicin, ethambutol, pyrazinamide, streptomycin, and thiacetazone) must be prescribed in an appropriate combination and in the right dosage, and they must be taken regularly under supervision and for a sufficient period of time. Treatment regimens with both isoniazid and rifampicin are the most effective regimens. Regimens can be shortened from 12-18 months to 6-8 months if they contain rifampicin. They are called short-course chemotherapy (SCC). SCC achieves higher completion rates and better cure rates than long-course chemotherapy. Health workers should abide by the national guidelines or, if there are no national guidelines, the World Health Organization's TB treatment guidelines. TB SCC begins with an initial intensive phase that lasts for a minimum of 2 months. It consists of a combination of at least 3 drugs to eliminate as many TB bacilli as possible and to prevent drug resistance. Fewer drugs are needed for the continuation phase that lasts for 4-6 months. Anti-TB drugs can be taken daily or 3 times a week. The final outcome of the intermittent therapy is the same as the daily therapy. Intermittent therapy is less suitable for young children. In children, the anti-TB regimen depends on pretreatment weight and is adjusted if necessary after 2 months. Health workers need to make sure that children with TB have no other infections. Malnutrition exacerbates the child's ability to fight infections. Young TB patients should be encouraged to eat. Adults are considered cured when their sputum smear changes from positive to negative; for children the best indications of a successful cure are completion of treatment, weight gain, and improvement in general health. Children rarely experience relapse and failure. Incorrect diagnosis or not taking the drugs usually accounts for failure. Sometimes drug resistant TB is responsible.     

Microbiology of tuberculosis

For a century, the diagnosis of tuberculosis, based on bacilloscopy and the isolation and identification of Mycobacterium tuberculosis in cultures, has been slow and not very sensitive. This has made it necessary on occasions to initiate treatment with tuberculostatics in an empirical way. The routine incorporation of liquid mediums and molecular genetic techniques in the final decade of the XX century brought an important advance by clearly increasing the sensitivity, precision and rapidity of diagnosis. The present blossoming of molecular techniques is making possible a better understanding of the disease's epidemiology, the factors of virulence and the mechanisms of resistance, which in the near future will give rise to new strategies of prevention and for treating the disease.     

结核病分类WS 196—2017

正前言本标准第3章为强制性条款,其余为推荐性条款。本标准按照GB/T1.1—2009给出的规则起草。本标准代替WS 196—2001《结核病分类》。本标准与WS 196—2001相比,主要技术变化如下:——增加了结核分枝杆菌潜伏感染者、非活动性结核病分类(见3.1、3.3);——将气管结核病、支气管结核病、结核性胸膜炎纳入肺结核分类和管理(见3.2.2.1)。