利用7H12B液体培养基快速鉴别人型和牛型结核分支杆菌

应用含ＴＣＨ１μｇ／ｍｌ的７Ｈ１２Ｂ培养基检测人型和牛型参比株各１株，１３种常见非结核分支杆菌参比杆各１株，临床示本检出的１５６株结核分支杆菌，１２株非结核分支杆菌。结果与含ＴＣＨ５μｇｍｌ罗－琼氏培养基鉴定结果完全一致，且所需时间明显综合，仅需２－４天。     

对硝基苯甲酸（PNB）替代BACTEC460NAP进行分支杆菌分群的初步研究

用ＰＮＢ替代ＢＡＣＴＥＣ系统中ＮＡＰ药物，将ＰＮＢ加入７Ｈ１２Ｂ培基内，鉴定结核菌与非结核分支杆菌两大菌群。实验１０株标准菌株和１１５株临床菌株；结果：人、牛型标准菌株在ＰＮＢ１２Ｂ１０μｇ／ｍｌ生长抑制。８株非结核标准菌株抑菌浓度ＰＮＢ〉５００μｇ／ｍｌ。以ＰＮＢ１００μｇ／ｍｌ１２Ｂ为界限，１１５株临床菌株ＰＮＢ与ＮＡＰ方法平行对照实验，鉴定结果：结核菌群符合率９７．８％，非结核菌群符合率９５     

鉴别人型及牛型结核分支杆菌的方法比较

本文对４４７株结核分支杆菌，用ＴＣＨ鉴别培养基、硝酸还原和烟酸试验３种方法进行人型及牛型结核分支杆菌的鉴别比较，３种方法的鉴别结果有明显统计学差异，（Ｐ＜０．０５～０．０１）。提示应用一种方法进行鉴别的准确性不可靠。因此，鉴别时，可使用ＴＣＨ鉴别培养基和烟酸试验两种方法，当结果不一致时应补作硝酸还原试验。这样可提高鉴别的可靠性。     

聚合酶链反应－单链构象多态性用于耐利福平结核分支杆菌rpoB基因突变的研究

目的研究结核分支杆菌ｒｐｏＢ基因突变及其与利福平（ＲＦＰ）耐药性的关系。方法以参考菌株结核分支杆菌Ｈ３７Ｒｖ为对照，用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）方法分析了４０株结核分支杆菌临床分离株。ＰＣＲ－ＳＳＣＰ方法由ＰＣＲ扩增和扩增产物ＤＮＡ单链多态性分析组成。结果两对引物ＰＣＲ扩增产物分别为４１１和２５８ｂｐ，其敏感性分别为５ｐｇ／μｌ、５００个菌／ｍｌ和１ｐｇ／μｌ、５００个菌／ｍｌ；均为属特异性。４０株结核分支杆菌临床分离株２５８ｂｐ扩增片段ＳＳＣＰ图谱的特点：以参考菌株结核分支杆菌Ｈ３７Ｒｖ为对照，１０株敏感株均无区别；单耐ＲＦＰ或包括ＲＦＰ多种抗结核药３０株，除３株外，其余２７株ＳＳＣＰ图谱有明显的区别；检测阳性率为９０％，特异性为１００％。结论ＰＣＲ－ＳＳＣＰ方法可检测出耐ＲＦＰ结核分支杆菌ｒｐｏＢ基因突变；该基因是ＲＦＰ的药物靶编码基因，它的突变与ＲＦＰ耐药性有密切关系，这有助于结核分支杆菌耐药性的快速检测和研究     

利用PCR方法直接从7H12B培养基中鉴定结核分支杆菌的探讨

以ＰＣＲ方法直接从７Ｈ１２Ｂ液体培养基中鉴别结核分支杆菌和非结核分支杆菌，与ＮＡＰ抑制试验对比，具有准确、快速，可应用于临床。     

采用16S rRNA序列分析法鉴定非结核分支杆菌

对第四次全国结核病流行病学抽样调查获得的经生化鉴定为非结核分支杆菌的 49株菌株进行了 16ＳｒＲＮＡ序列分析法鉴定 ,并与生化法进行了比较。材料与方法　菌株 :牛分支杆菌 (ＡＴＣＣ 192 10 ) ,偶然分支杆菌 (ＡＴＣＣ6841) ,标准菌株胞内分支杆菌 (ＡＴＣＣ 3 5 772 ) ,不产色分支杆菌 (ＤＳＭ 44 164 ) ,瘰疬分支杆菌 (ＤＳＭ 43 992 )均由国家结核病参比实验室提供。获取方法 :参照《第四次全国结核病流行病学抽样调查工作手册 (修订本 )》。聚合酶链反应 (ＰＣＲ)扩增 16ＳｒＲＮＡ的片段 :取罗氏培养基培养的非结核分支杆菌 ,灭活…     

结核分支杆菌重组38000蛋白抗原诱导豚鼠迟发性超敏？…

探讨结核分支杆菌重组３８００抗原诱导豚鼠产生迟发性超敏反应的能力及影响因素。方法常规皮肤试验法：Ｈ３７Ｒｖ致敏豚鼠,皮试时以生理盐水为阻对照,５ＩＵＰＰＤ为阳性对照,０．１μｇ、０．２μｇ、０．４μｇｒ几ＵＸＥ ＵＫ ＨＣ ＭＷ ＩＹ ＴＭＤＦ,ＩＹ     

氧氟沙星和环丙沙星对快速生长分支杆菌最低抑菌浓度测定

以Ｂａｃｔｅｃ４６０ＴＢ检测议测定了氧氟沙星（ＯＦＬＸ）和环丙沙星（ＣＰＦＸ）对１１株快速生长（速生）分支杆菌、３株标准菌株的最低抑菌浓度（ＭＩＣ＿９９），结果两药对龟型分支杆菌龟亚种ＭＩＣ＿９９最高，分别为１．０和１．０～２．０μｇ／ｍｌ，对龟型分支杆菌脓肿亚种及偶发分支杆菌ＭＩＣ＿９９相近，分别为０．２５和０．５～１．０μｇ／ｍｌ，对１株转黄和１株待定型速生分支杆菌ＭＩＣ＿９９分别为≤０．１２５，０．５和≤０．１２５，１．０μｇ／ｍｌ。结果表明，体外两药对速生分支杆菌有强抑菌作用，值得进行临床验证。     

利用PCR方法直接从7H12B培养基中鉴定结核分支杆菌的探讨

以ＰＣＲ方法直接从７Ｈ１２Ｂ液体培养基中鉴别结核分支杆菌和非结核分支杆菌，与ＮＡＰ抑制试验对比，具有准确、快速，可应用于临床。     

分支杆菌鉴定方法的比较研究

比较了微菌落法、多重聚合酶链反应 (ＰＣＲ)法和传统法用于分支杆菌临床分离物菌种鉴定的效果 ,现报告如下。材料与方法 :试验菌株 :(1)标准菌株 :结核分支杆菌Ｈ37ＲＶ、牛分支杆菌、鸟分支杆菌、胞内分支杆菌、堪萨斯分支杆菌、瘰疬分支杆菌、偶然分支杆菌和耻垢分支杆菌标准株 ,均购自北京结核病胸部肿瘤研究所。 (2 )临床分离株 :来自我院结核病住院患者的痰、胸液和脑脊液。传统分型鉴定试验 :(1)培养鉴定试验 :按常规制备对硝基苯甲酸 (ＰＮＢ)和噻吩 2 羧酸肼 (ＴＣＨ)培养基 ,取试验菌株菌悬液 (10 -2 /ｍｌ) 0 1ｍｌ接种 ,37℃…     

噬菌体生物扩增法与其它方法在结核菌检测中的比较

目的通过三种方法的比较,研究噬菌体生物扩增法在结核菌检测中的意义和可行性。方法同一份标本分别做抗酸杆菌涂片法、BACTEC MGIT 960培养法和噬菌体生物扩增法。结果检测的243例标本中,噬菌体生物扩增法阳性104例,抗酸杆菌涂片法阳性44例,BACTEC MGIT 960培养法95例阳性。结论噬菌体生物扩增法检测结核菌阳性率远高于涂片法,略高于BACTEC MGIT 960培养法。噬菌体生物扩增法可以快速鉴定结核分枝杆菌,用其检测标本中的结核分枝杆菌具有很高的特异性和较高的敏感性。     

Variable agreement among experts regarding Mycobacterium avium complex lung disease

Data regarding many clinical aspects of pulmonary Mycobacterium avium complex (pMAC) are lacking. Guidelines rely substantially upon expert opinion, integrated through face‐to‐face meetings, variably weighting individual opinions. We surveyed North American non‐tuberculous mycobacteria experts regarding clinical aspects of pMAC using Delphi methods. Nineteen of 26 invited experts (73%) responded, with extensive variability. Convergence could not be reached for most questions. Respondents described extensive uncertainty around specific issues. Findings underscore urgent need for more research.     

青浦区552株结核分枝杆菌原发耐药性分析

目的了解青浦区1999年～2008年新登记肺结核患者的原发耐药情况。方法采用绝对浓度法对新发初治肺结核患者痰标本培养菌株进行链霉素（S）、异烟肼（H）、利福平（R）和乙胺丁醇（E）4种抗结核药物的耐药性测定，统计分析原发耐药情况。结果原发总耐药率由1999年～2000年的9．2％上升至2007年～2008年的15．5％；S、H、E的耐药率由1999年的5．0％、7．1％、2．8％上升至2008年的12．1％、13．8％、3．4％；R的耐药率由1999年的5．0％下降至2007年～2008年的4．3％；原发耐多药率（MDR．TB）平均为4．5％。2药、3药的耐药率分别由1999年的2．1％、0％上升至2008年的6．0％、1．8％。1药、4药的耐药率基本在同一水平。结论青浦区肺结核患者临床分离株原发耐药性成上升趋势，耐多药菌株仍占一定比例。     

120例矽肺结核患者结核分枝杆菌L型致病情况研究

目的探讨研究120例矽肺结核患者结核分枝杆菌L型致病情况的价值。方法对矽肺结核组和矽肺患者，分别进行痰结核分枝杆菌和痰结核分枝杆菌L型培养，并对结果进行对照观察。结果矽肺结核120例痰结核分枝杆菌培养阳性14例，阳性率11．6％，同时出现结核分枝杆菌L型培养阳性10例，占71．4％；痰结核分枝杆菌L型培养阳性58例，阳性率48％（P〈0．01）。结论结核分枝杆菌L型培养方法方便快速，能提高结核分枝杆菌的检出率，对矽肺结核的早期诊断及减少矽肺结核复发的漏、误诊有肯定的价值。     

An adenosine triphosphate-independent proteasome activator contributes to the virulence of Mycobacterium tuberculosis

Mycobacterium tuberculosis encodes a proteasome that is highly similar to eukaryotic proteasomes and is required to cause lethal infections in animals. The only pathway known to target proteins for proteasomal degradation in bacteria is pupylation, which is functionally analogous to eukaryotic ubiquitylation. However, evidence suggests that the M. tuberculosis proteasome contributes to pupylation-independent pathways as well. To identify new proteasome cofactors that might contribute to such pathways, we isolated proteins that bound to proteasomes overproduced in M. tuberculosis and found a previously uncharacterized protein, Rv3780, which formed rings and capped M. tuberculosis proteasome core particles. Rv3780 enhanced peptide and protein degradation by proteasomes in an adenosine triphosphate (ATP)-independent manner. We identified putative Rv3780-dependent proteasome substrates and found that Rv3780 promoted robust degradation of the heat shock protein repressor, HspR. Importantly, an M. tuberculosis Rv3780 mutant had a general growth defect, was sensitive to heat stress, and was attenuated for growth in mice. Collectively, these data demonstrate that ATP-independent proteasome activators are not confined to eukaryotes and can contribute to the virulence of one the world''s most devastating pathogens.Proteasomes are compartmentalized proteases essential for the viability of all eukaryotes and archaea. The proteolytically active component of the proteasome is the 20S core particle (CP), a ∼700-kDa complex composed of four heptameric rings stacked axially to form a barrel (reviewed in refs. 1 and 2). The active site of the 20S CP is buried deep within its core, rendering it inaccessible to folded proteins (3, 4). Further protection of the active site is afforded by “gating” amino acids that block the entrance to the proteasome core. Thus, proteasomal degradation is critically dependent on proteasome activators, which bind to either end of the 20S CP to reposition its gating residues and allow access to doomed substrates (reviewed in ref. 5).The best-characterized proteasome activator is the eukaryotic 19S regulatory particle (RP). The 19S RP is a ∼700-kDa heteromeric complex that binds either or both ends of the 20S CP to form the 26S proteasome, which is responsible for degrading proteins in an adenosine triphosphate (ATP)-dependent manner (6–8). The unfolding activities of the 19S RP come from its base, a heterohexameric ring of ATPases associated with diverse cellular activities (AAA ATPases) that directly bind to the 20S CP (reviewed in ref. 2). Ubiquitylation, which is the posttranslational modification of doomed proteins with the small protein ubiquitin, is the major signal that targets proteins for degradation by the 26S proteasome. Because ubiquitylation is an essential regulator of virtually every cellular pathway in eukaryotes, a considerable amount of work has focused on the biochemistry and biology of the 26S proteasome, leading to the identification of many substrates and the reconstitution of robust in vitro degradation (reviewed in refs. 2 and 9).In addition to the 19S RP, there are several cofactors that activate proteasome activity without ATP. These include Blm10/PA200 (10, 11), archaeal PbaB (12), and the 11S activators. The 11S activators are a family of small proteins that form heptameric rings and enhance peptide degradation upon binding to 20S CPs. Simple eukaryotes encode a single 11S activator, PA26 (13), whereas higher eukaryotes encode three 11S activator isoforms: PA28-α, -β, and -γ (14–16). An abundance of work has detailed the biochemical characteristics of the 11S activators, which has been crucial in understanding the structural basis of proteasomal gate opening (17–21). However, their precise biological roles remain controversial. A multitude of studies have demonstrated that PA28-α/β alter the peptide products of proteasomal degradation to influence major histocompatibility complex class I antigen presentation (22, 23). However, PA28-α/β–deficient mice are not remarkably defective in antigen presentation or immunity (24). Similarly, only a handful of PA28-γ–dependent protein substrates have been described, and few studies have demonstrated a significant physiologic role for PA28-γ in mammals (25–29). As a result, our understanding of the biological relevance of eukaryotic ATP-independent activators has lagged behind that of the 19S RP.Although the bulk of research into proteasomes has been in eukaryotes, they are also present in bacteria of the orders Nitrospirales and Actinomycetales, which includes the human pathogen Mycobacterium tuberculosis. M. tuberculosis infects one-third of the world’s population, killing >1.5 million people each year (www.who.int/mediacentre/factsheets/fs104/en/). There is an urgent need for the identification of novel antitubercular drug targets because resistance to currently available drugs is on the rise. One potential target is the prokaryotic ubiquitin-like protein (Pup)-proteasome system (PPS), a pathway used by M. tuberculosis and other proteasome-bearing bacteria to target proteins for degradation. The PPS centers on the small protein modifier Pup (30). The C terminus of Pup is covalently attached to a lysine of a target protein by the Pup ligase proteasome accessory factor A (PafA) (31) and is then bound by the homohexameric AAA ATPase mycobacterial proteasome ATPase (Mpa), which resembles the Rpt ATPases found in the bases of eukaryotic 19S RPs (32–35). Mpa unfolds pupylated proteins to be degraded by the M. tuberculosis 20S CP, which is structurally similar to the eukaryotic 20S CP but is composed of just two unique subunits, PrcA and PrcB (36, 37). Because the PPS is chemically distinct from the eukaryotic ubiquitin–proteasome pathway, and because proteasomal degradation of pupylated proteins is required for full M. tuberculosis virulence (32, 38–40), the PPS represents an attractive therapeutic target for tuberculosis.To date, pupylation is the only pathway known to target proteins for proteasomal degradation in bacteria. However, several studies suggest that the 20S CP may have pupylation-independent functions, because chemical inhibition of the 20S CP or genetic deletion of the 20S CP genes, but not mpa or genes involved in pupylation, produces a severe growth defect in liquid culture and on solid medium (38–41). Furthermore, although mpa and pupylation mutants are severely attenuated for growth in mice, they are able to persist for the natural lifespan of infected animals; in contrast, 20S CP mutants fail to persist in mice (40, 41). It is therefore likely that the M. tuberculosis proteasome has Pup-independent functions.In an effort to identify proteasomal cofactors in M. tuberculosis that might contribute to new degradation pathways, we used a catalytically inactive proteasome trap to stabilize normally transient interactions of the M. tuberculosis 20S CP. The proteasome trap interacted with Rv3780, a previously uncharacterized protein. Rv3780 formed oligomeric rings and bound to M. tuberculosis 20S CPs. This interaction required a conserved C-terminal motif in Rv3780, reminiscent of what has been described for eukaryotic proteasome activators. We show that Rv3780 promoted the robust ATP-independent proteasomal degradation of short peptides, an unfolded protein, and a native M. tuberculosis protein in vitro. Importantly, Rv3780 contributed to M. tuberculosis resistance to heat stress and virulence in a mouse infection model.     

母牛分枝杆菌菌苗对复治性肺结核的辅助治疗作用

目的观察母牛分枝杆菌菌苗（微卡）在复治性肺结核治疗中的疗效。方法将146例痰抗酸杆菌（AFB）阳性复治肺结核患者分成微卡治疗组（n：90）和对照组（n=56）疗程各6个月，前者采用复治化疗方案加用微卡治疗，后者仪用复治方案治疗。结果疗程结束时微卡治疗组和对照组痰阴转率分别为85．56％（77／90）、67．86％（38／56），差异有显著性（P〈0．05）。微卡治疗组和对照组有效率分别为81．11％（73／90）、44．64％（25／56），两组比较差异有显著性（P〈0．001）。结论微卡能改善难治性肺结核患者的细胞免疫功能，有助于痰菌阴转和病灶吸收，可作为复治性肺结核化疗的辅助药物。     

微卡辅助治疗复治涂阳肺结核的临床效果

目的评价微卡（母牛分枝杆菌菌苗）作为免疫调节剂辅助治疗复治涂阳肺结核的疗效。方法将80例复治涂阳肺结核病人随机分为治疗组40例及对照组40例，两组化疗方案相同3SHRZE／6HR，治疗组肌肉注射微卡，每半月1次，共9个月，观察药物不良反应。结果疗程结束时，治疗组痰菌阴转率为92．5％，明显高于对照组77．5％（P〈0．01）；治疗组病灶吸收，空洞关闭率分别为97．5％和91．7％均明显高于对照组85．0％和78．3％（P〈0．01），有统计学意义。结论微卡是一种新型免疫调节剂，可提高复治涂阳肺结核的疗效，值得推广。     

硝酸盐还原试验快速检测结核分枝杆菌药敏试验

目的探讨一种廉价、简便、快速的检测结核分枝杆菌药敏的方法。方法以绝对浓度法对照,对92株结核分枝杆菌进行硝酸盐还原酶试验,并比较两种方法。结果四种一线抗痨药物利福平、异烟肼、链霉素、乙胺丁醇的敏感度分别为96.83%、97.22%、94.11%、88.89%;特异度为92.59%、85.0%、87.80%、96.43%;符合率为93.48%、96.74%、91.30%、93.48%。两种方法无显著性差异,但硝酸盐还原酶法的检测时间要比绝对浓度法快2～3周。结论硝酸盐还原酶法是一种快速、准确检测结核药敏的方法。     

艾滋病合并鸟-胞内分枝杆菌复合群 中枢神经系统感染1例并文献复习

[摘要]　非结核分枝杆菌相关颅内感染相对罕见。本文报道了1例艾滋病合并鸟-胞内分枝杆菌复合群中枢系统感染病例。病例系1名45岁女性患者，因“头晕3月，加重伴头痛、失语1周”住院，入院后进行脑脊液检查，并利用二代宏基因组测序发现感染病原菌为鸟-胞内分枝杆菌复合群，从而采取相应的治疗措施，最终获得治愈。本文通过复习有关资料，深入探讨了艾滋病和非结核分枝杆菌相关性颅内感染的临床表现和处理，提示临床医生警惕非结核分枝杆菌相关性颅内感染及其严重性，及时确定病因并实施相应的干预措施。