结核抗体蛋白芯片法对老年肺结核的诊断价值研究

目的探讨结核抗体蛋白芯片法对老年肺结核的诊断价值。方法选择93例肺结核及77例肺部非特异性感染老年患者,均行痰涂片抗酸染色试验,均采用结核分枝杆菌IgG抗体检测试剂盒(蛋白芯片)检测抗结核分枝杆菌脂阿拉伯甘露醇(LAM)、基因工程重组的16 kD和38 kD的IgG抗体,计算两种方法诊断结核分枝杆菌感染的敏感性及特异性;并分别构建受试者工作特征曲线,通过比较曲线下面积(area under the curve,AUC)评价其诊断效能。结果蛋白芯片法和痰涂片抗酸染色试验诊断结核分枝杆菌感染的敏感性分别为68.82%和33.33%,特异性分别为100.00%和97.40%;AUC分别为0.84和0.65,提示蛋白芯片法的诊断效能显著高于痰涂片抗酸染色法,差异有统计学意义(P0.01)。结论蛋白芯片法检测抗结核分枝杆菌LAM、16 kD和38 kD抗体快捷简便,对诊断老年肺结核有较高的实用价值。     

蛋白芯片法与金标法及抗酸染色法诊断结核病的比较

目的将诊断结核病的蛋白芯片法与金标法及抗酸染色法进行比较,以评价蛋白芯片的临床应用价值。方法用蛋白芯片法、金标法及抗酸染色法检测75例确诊结核病人、30例肺部其它疾病患者和健康体检者的血清标本,并进行相关统计分析。结果蛋白芯片法、金标法及抗酸染色法检出结核分枝杆菌的灵敏度为64%,72%及22.7%,前两者比较无显著性差异(P>0.05),但均高于抗酸染色法(P<0.05)。三者的特异性分别为90%,76.7%及96.7%。结论蛋白芯片法检测结核分枝杆菌,与金标法及抗酸染色法比较,具有简便、快速、敏感性较高和特异性强等优点,是临床辅助诊断结核病的有效方法。     

结核蛋白芯片检测诊断结核病的价值

目的探讨结核蛋白芯片检测诊断结核病的价值。方法使用结核蛋白芯片系统检测4 093例患者的血清结核抗体,其中结核病441例,非结核病3 652例。采用结核蛋白芯片技术检测结核分枝杆菌中ESAT-6、CFP10、LAM、38KD和16KD 5种成分,分析其结果。结果 441例结核患者中297例结核抗体阳性,3 652例非结核患者中,647例阳性,结核蛋白芯片检测的灵敏度为67.35%,特异度为82.28%。结论结核蛋白芯片是诊断结核病的较有效方法,具有特异性好、快速方便等优点,对快速诊断结核病起辅助作用。     

结核分枝杆菌γ-干扰素体外释放定量试验对结核病诊断价值的回顾性分析

摘要：目的：回顾性分析结核分枝杆菌γ-干扰素体外释放定量试验(TB-IGRA)对结核病的诊断价值。 方法：收集2013年10月至2014年9月进行TB-IGRA检测的住院患者367例,其中结核病患者89例,包括69例肺结核患者和20例肺外结核病患者,非结核疾病患者278例作为对照,分析TB-IGRA法诊断结核病的敏感性、特异性、阳性预测值和阴性预测值,并与痰涂片镜检法、结核抗体胶体金法、结核抗体蛋白芯片法进行比较。 结果：TB-IGRA在肺结核和肺外结核患者中检出的阳性率分别为86.9%和95.0%,差异无统计学意义(P>0.05),且两组间IFN-γ释放水平也无统计学意义（P>0.05)。而结核病组的IFN-γ水平明显高于非结核疾病组（P<0.01）,且治疗有效时IFN-γ水平显著降低甚至转阴。TB-IGRA法的诊断敏感性、特异性、阳性预测值、阴性预测值分别为88.8%、76.6%、54.8%和95.5%。痰涂片镜检、胶体金法检测结核抗体和蛋白质芯片法检测结核抗体的敏感性分别为32.5%、13.7%和38.5%。TB IGRA与另3种方法敏感性差异有统计学意义(P<0.01)。 结论：TB-IGRA对结核病具有良好的敏感性和阴性预测值,是一种有价值的结核病辅助诊断方法,但特异性和阳性预测值较低,阳性结果还需结合临床加以判断。动态监测IFN-γ水平可以提示抗结核治疗疗效。     

结核生物蛋白芯片技术在结核病临床诊断中的价值探讨

目的利用结核生物芯片技术,探讨国产重组结核分枝杆菌蛋白38kD(γTPA38)、16kD(γTPA16)和脂阿拉伯甘露糖(LAM)用于肺结核与肺外结核的早期诊断价值。方法采用CCD原理结合视频采集方法,应用结核生物芯片检测系统,对门诊及住院患者血清标本3448例进行了检测。结果 LAM、16kD、38kD结核生物蛋白芯片联合检测门诊患者782例,阳性219例,阳性率为28.0%(219/782)。住院患者2666例,结核生物蛋白芯片联合检测阳性1368例,阳性率为52.0%(1386/2666)。实验室检查了结核生物蛋白芯片联合检测阳性的1386例住院患者的细菌涂片,其中菌阳患者213例,菌阴患者1155例。还对69例健康人群及肺部其他疾病患者血清标本进行了16kD、38kD、LAM抗体的检测,结果阴性66例,特异性为95.7%(66/69),3448例患者3种抗原联合检测,结果阳性1587例,敏感度为46.0%(1587/3448)。结论结核生物芯片检测系统联合运用16kD、38kD、LAM3种抗原检测其对应的抗体有较好的灵敏度和很高的特异性。对结核病血清学诊断有较高的参考价值。     

不同免疫学检验方法对于结核病的临床价值研究

目的探讨蛋白芯片法与胶体金法对于结核病的临床价值。方法选取该院从2015年2~11月所收治的90例确诊结核病患者作为临床研究对象,同时采用蛋白芯片法与胶体金法进行检测并比较两种检测方法的差异性,分析不同方法在结核病辅助诊断当中的临床价值。结果结核组患者的胶体金法阳性反应率为73.3%(66/90),显著高于对照组[7.8%(7/90)],差异有统计学意义(P0.05);结核组患者蛋白芯片法的阿拉伯甘露糖脂(LAM)、蛋白16kD和蛋白38kD阳性反应率为53.3%(48/90)、22.2%(20/90)和53.3(48/90),高于对照组5.6%(5/90)、0%(0/60)和5.6%(5/90),差异有统计学意义(P0.05)。结果表明,结核组患者的胶体金法阳性反应率显著高于蛋白芯片法阳性反应率,差异有统计学意义(P0.05)。结论胶体金法在结核病的辅助诊断中更有优势,其灵敏度高、操作简单,有利于推广,对于结核病的早诊断、早治疗具有较强实用性,是一种比较理想的免疫学检验方法。值得在今后临床工作中推广应用。     

结核蛋白芯片对肺结核及肺外结核的诊断价值

目的探讨结核蛋白芯片检测对诊断肺结核及肺外结核的价值。方法使用蛋白芯片系统检测血清结核抗体的病例共4765例,其中肺结核334例,肺外结核83例,肺结核并肺外结核69例,非结核病4279例。我们采用的蛋白芯片包含了结核分枝杆菌（MTB）中5种成分,即ESAT-6、CFP10、LAM、38KD和16KD。结果 334例肺结核中227例结核抗体阳性,83例肺外结核中50例阳性,69例肺结核并肺外结核中51例阳性,4279例非结核病797例阳性,结核蛋白芯片检测肺结核的阳性率为67.96%,肺外结核的阳性率为60.24%,肺结核并肺外结核的阳性率为73.91%,非结核病的阳性率为18.62%。结论结核蛋白芯片是对肺结核及肺外结核的辅助诊断有重要价值。但在淋巴结结核、结核性胸膜炎中阳性率偏低,敏感性稍差。     

结核生物蛋白芯片技术在临床诊断中的意义

目的 利用芯片技术,建立结核LAM,16-kDa,38-kDa蛋白芯片检测结核分技杆菌的方法,以期为结核与肺外结核的早期诊断提供参考.方法 采用CCD原理结合视频采集方法,应用结棱蛋白膜芯片进行1 623例检测.结果 LAM,16-kDa,38-kDa蛋白芯片联合检测住院病人阳性率53.2%(396/744);门诊病人30.0%(264/879).结论 结核多种抗原的蛋白芯片检测系统联合运用三种抗原联合检测对结核病的诊治有积极的意义.     

结核蛋白芯片在结核病辅助诊断中的应用

目的了解结核蛋白芯片检测在结核病早期快速诊断中的应用价值。方法收集2008～2010年门诊和住院确诊的结核患者156例(痰菌阳性42例),非结核患者40例。通过结核芯片系统检测3种抗体,任意一种抗体阳性,结果判定为阳性。结果单一抗体阳性率为33.33%,两种抗体阳性率为12.82%,3种抗体阳性率为21.79%,所有抗体总阳性率为67.95%,特异性约为90.00%;痰涂片抗酸染色阳性率为26.92%,特异性为97.50%。结论结核蛋白芯片检测法具有简便、快速、敏感性高、特异性强的优点,能早期、快速诊断结核病,是临床辅助诊断结核病的一种有效方法。     

结核抗体对老年结核病患者的诊断价值探讨

目的评价结核抗体对老年结核病患者的诊断价值。方法选择老年住院患者148例,其中确诊结核病38例,采用酶联免疫吸附试验（ELISA）检测其结核抗体,用临床流行病学方法分别计算结核抗体检测对结核病诊断的敏感性、特异性、似然比、预测值及不同患病率下的验后概率。结果结核抗体检测对结核病诊断的敏感性和特异性分别为55.26%和90.00%,阳性和阴性似然比分别为5.526和0.497,阳性预测值为65.63%,阴性预测值为85.34%。患病率为25.68%时,验后概率为65.64%。结论结核抗体检测对老年结核病患者的诊断价值有限,临床医师应谨慎对待结核抗体检测阳性结果,其诊断价值与所处医院患者结核病患病率有关。     

DNA 微阵列芯片法用于检测结核分枝杆菌耐药性的研究

目的：通过与传统的比例法在结核分枝杆菌耐药性检测的比较,评价 DNA 微阵列法用于检测结核分枝杆菌耐药性的可行性。方法随机抽取本院2012年1月至2013年3月从临床标本中分离培养所得的结核分枝杆菌54株,通过 DNA 微阵列法和比例法分别进行异烟肼和利福平的耐药性检测并对结果进行比较分析。结果以比例法为金标准,DNA 微阵列法对异烟肼和利福平的耐药检测结果与比例法的符合率分别为75．0％、91．0％。结论 DNA 微阵列技术适用于临床一线耐药结核分枝杆菌的快速筛查。     

DNA微阵列芯片法与常规生化法在分枝杆菌菌种鉴定方面的比较

目的通过DNA微阵列芯片法进行分枝杆菌菌种鉴定与常规生化法进行比较,分析其特点,提高分枝杆菌菌种鉴定水平,更好的为临床服务。方法选择我院2010年1月至2013年3月从临床标本中分离培养后所得的结核分枝杆菌复合群12株（其中含1株牛型结核分枝杆菌）,非结核分枝杆菌3l株,分别用DNA微阵列芯片法和常规生化法进行鉴定。结果DNA微阵列芯片法进行分枝杆菌菌型鉴定与普通生化培养法鉴定结果符合率为100％,对常规生化法未定型的两株非结核分枝杆菌也分别定型为1株土分枝杆菌,1株耻垢分枝杆菌。结论DNA微阵列芯片法与常规生化法相比在分枝杆菌菌型鉴定中具有更快速、准确的特点,是分枝杆菌菌型鉴定的有利工具。     

DNA微阵列芯片法在海南地区结核病诊断及耐药性检测中的应用

目的 探讨DNA微阵列芯片法在海南地区结核病诊断及耐药性检测中的应用.方法 采用抗酸杆菌涂片法、罗氏培养法、比例法药敏试验及DNA微阵列芯片法对海南地区的2069例疑似结核病患者痰标本进行检测,并对结核分枝杆菌检出率、耐药性及耐药基因突变特征进行分析.结果 DNA微阵列芯片法检测结核分枝杆菌检出率为明显高于罗氏培养法和...     

Detection of rifampin-resistant Mycobacterium tuberculosis strains by using a specialized oligonucleotide microarray

DNA microarray represents one of the major advances in diagnostic sequencing of polymerase chain reaction (PCR) products. Until now, arrays have been relatively expensive, complex to perform, and difficult to interpret, limiting their wide application in the clinical laboratory. A moderate-density oligonucleotide microarray that can rapidly identify Mycobacterium tuberculosis rifampin-resistant strains was developed. The method is based on the detection of point mutations and other rearrangements in the rpoB gene region determining rifampin resistance. Rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Fifty-three rifampin-resistant M. tuberculosis and 15 rifampin-susceptible M. tuberculosis were tested and results were concordant with those based on culture drug susceptibility testing and sequencing. Rifampin-resistant clinical isolates were detected in as little as 1.5 hours after PCR amplification with visual results. It is demonstrated that oligonucleotide microarray is an efficient, specialized technique to implement and can be used as a rapid method for detecting rifampin resistance to complement standard culture-based method.     

Low penetrance,broad resistance,and favorable outcome of interleukin 12 receptor beta1 deficiency: medical and immunological implications

The clinical phenotype of interleukin 12 receptor beta1 chain (IL-12Rbeta1) deficiency and the function of human IL-12 in host defense remain largely unknown, due to the small number of patients reported. We now report 41 patients with complete IL-12Rbeta1 deficiency from 17 countries. The only opportunistic infections observed, in 34 patients, were of childhood onset and caused by weakly virulent Salmonella or Mycobacteria (Bacille Calmette-Guérin -BCG- and environmental Mycobacteria). Three patients had clinical tuberculosis, one of whom also had salmonellosis. Unlike salmonellosis, mycobacterial infections did not recur. BCG inoculation and BCG disease were both effective against subsequent environmental mycobacteriosis, but not against salmonellosis. Excluding the probands, seven of the 12 affected siblings have remained free of case-definition opportunistic infection. Finally, only five deaths occurred in childhood, and the remaining 36 patients are alive and well. Thus, a diagnosis of IL-12Rbeta1 deficiency should be considered in children with opportunistic mycobacteriosis or salmonellosis; healthy siblings of probands and selected cases of tuberculosis should also be investigated. The overall prognosis is good due to broad resistance to infection and the low penetrance and favorable outcome of infections. Unexpectedly, human IL-12 is redundant in protective immunity against most microorganisms other than Mycobacteria and Salmonella. Moreover, IL-12 is redundant for primary immunity to Mycobacteria and Salmonella in many individuals and for secondary immunity to Mycobacteria but not to Salmonella in most.     

Five-antituberculosis Drug-resistance Genes Detection Using Array System

Detection of resistance to drugs for Mycobacterium tuberculosis takes about two months from the sample collection using culture-based methods. To test a rapid method for detection of resistance for five antituberculosis drugs using DNA microarray and to examine its potential for clinical use, we employed a DNA microarray for detection of seven mutations genes related to resistance of five kinds of antituberculous drugs using Mycobacterium tuberculosis DNA isolated from sputum. The results of microarray analysis were compared with the results of a standard culture method of Lowenstein-jensen drug sensitivity testing system. DNA microarray analysis showed a high sensitivity (>90%) for all five drugs. Specificity of rifampicin and ethambutol were nearly 90%, however specificity of isoniazid (60%) and kanamycin (67%) were not enough. The amount of Mycobacterium tuberculosis DNA required for microarray analysis corresponded to at least 1-9 Acid-Fast Bacilli per 10 fields by carbolfuchsin staining. DNA microarray analysis appears to be useful for estimation of drug resistances, nevertheless its limitations. To minimize misunderstanding, it is necessary to confirm the number of bacilli in the sputum, and culture method is needed for comparison when use the PCR-based array system.     

噬菌体生物扩增法环节中最佳感染时间的探讨

目的 探讨影响噬菌体生物扩增法检出结核分枝杆菌敏感性的关键环节,即最佳感染时间.方法 观察噬菌体生物扩增法中噬菌体D29分别感染结核分枝杆菌0、0.5、1、1.5、2、3、4、5、6和8 h对噬菌斑数量的影响.并且在电镜下直接观察噬菌体D29与结核分枝杆菌和耻垢分枝杆菌中共同孵育0、15、30、60、90、120、180、240、360 min时,噬菌体的吸附、感染和增殖情况.综合上述两种方法 结果 确定噬菌体生物扩增法环节中的最佳感染时间.结果 噬菌体感染结核分枝杆菌3～4 h时,噬菌斑形成数量达到高峰[(321±2.65)PFU],4 h后噬菌斑开始出现下降,5 h时噬菌斑数为(300±5.62)PFU.在电镜下直接观察到噬菌体D29与耻垢分枝杆菌接触15 min后,即开始吸附耻垢分枝杆菌,3 h后完成子代噬菌体的合成.噬菌体D29与结核分枝杆菌接触3～4 h时吸附感染部分结核分枝杆菌,6 h后完成子代噬菌体合成.结论 采用噬菌体生物扩增法检测标本中结核分枝杆菌需要多个环节,并且可以同时检测大量标本,为提高检测的敏感性以3 h作为感染时间最为恰当.     

海南省肺部感染患者非结核分枝杆菌的菌种鉴定分析

目的 通过对海南省肺部感染患者非结核分枝杆菌（NTM）进行菌种鉴定,探讨海南省NTM肺部感染菌种的类型及人群分布情况。方法 收集2016年7月至2021年6月期间就诊于海南医学院第二附属医院疑似肺结核患者的呼吸道标本进行分枝杆菌培养,对培养阳性标本进行对硝基苯甲酸（PNB）/噻吩-2-羧酸肼（TCH）培养菌型初步鉴定,采用DNA微阵列芯片技术进行分枝杆菌菌种鉴定,无法确定菌种的菌株进一步采用基因测序法鉴定。结果 共收集8 507例疑似肺结核患者的呼吸道标本,剔除重复病例后,有318例经PNB/TCH培养初步鉴定为NTM,315例经DNA微阵列芯片技术和热休克蛋白65(hsp65)基因测序鉴定为NTM。其中308例患者为单一感染模式,6例MTB+NTM和1例2种不同NTM的混合感染模式。快速生长分枝杆菌128株占40.5%,以龟/脓肿分枝杆菌为主（占32.9%）;缓慢生长分枝杆菌188株占59.5%,以胞内分枝杆菌为主（占39.6%）。女性患者多于男性且好发于中老年人,男女性别比为0.79∶1。男女性患者年龄分布差异有统计学意义（Z=2.200,P<0.05）,>40岁的患者...     

Lupus autoantibodies target ribosomal P proteins

All nine SLE (systemic lupus erythematosus) sera with antiribosomal antibody activity targeted the same three ribosomal protein antigens, of molecular masses 38 and 17/19 kD when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. One serum reacted with an additional protein of approximately kD. Ribosomal subunit fractionation by composite gel electrophoresis and sucrose density ultracentrifugation showed that these proteins were part of the large subunit. Isoelectric focusing in agarose, and two-dimensional polyacrylamide gel electrophoresis revealed that the antigens had pI between 4.5 and 6.5, but that the 17/19 kD antigens were more acidic than the 38 kD antigen. Similarities in the molecular masses, charges, as well as the presence of highly conserved crossreactive epitopes, failure to bind to carboxymethylcellulose at pH 4.2, and extractability of the 17/19 kD proteins by 400 mM NH4Cl-ethanol at 0 degrees C indicated that these antigens were analogous to the proteins P0 (38 kD) and P1/P2 (17/19 kD) described previously (25, 36). Co-identity was confirmed using reference antibodies and antigen. Although antibodies to these proteins were only found in 5-10% of more than 50 sera screened by radioimmunoassay or Western blotting, the selective production of antibodies to epitopes on three (out of a total of more than 80) ribosomal proteins may provide further clues to autoantibody induction of SLE.