显微镜检查和血清学诊断相结合筛查活动性结核

地点:美国加洲大学圣迭戈医学中心.目标:建立一种包括抗体检测的结核病(TB)简单筛查策略,以提高痰抗酸杆菌镜检(AFB涂片)的准确性.方法:血清标本来自190名可疑活动性结核患者.结核病的诊断通过结核杆菌培养确定.HIV感染状况由商业化的血清学试验确定.IgG抗体水平使用纯化的结核杆菌抗原用ELISA法测定.从130名随机选择的病人资料用于制定筛查策略,另60名病人资料用于验证.结果:抗酸菌涂片敏感性为70%,特异性为88%.单个或多个抗原的ELISA同AFB涂片及HIV状况相结合诊断模式的敏感性比每个单项试验都高.包括4种抗原ELISA(38KD抗原,脂阿拉伯甘露聚糖,MPT-64和谷氨酰胺合成酶)诊断敏感性为93%,特异性为76%.和AFB涂片相比其敏感性较高,而特异性无统计学差异.结论:本研究建议建立将多种抗原ELISA同AFB涂片和HIV检查整合在一起的筛查策略.     

三种血清学方法检测抗结核抗体对肺结核诊断价值的评估

目前，肺结核辅助诊断方法主要包括胸部X线、痰细菌学及血清学检查。胸部X线检查缺乏特异性；痰涂片抗酸染色和结桉菌培养阳性率低；近年来迅速发展的分子生物学技术如聚合酶链反应(PCR)，所需设备复杂，实验费用昂贵，假阳性率高。因此结核病血清学诊断方法日益受到重视，并且有了很大进展。献报导，用MycoDot快速诊断试剂盒检测血清中脂阿拉怕甘露糖(LAM)-IgG抗体可诊断活动性结核病，用38KDa抗原快速检测血清中抗结核抗体可诊断肺结核。     

1983-2001年阿根廷结核病实验室网络抗酸杆菌涂片显微镜检查的质量评价

地点阿根廷结核病实验室网络.目标(1)评价1983-2001年间督导的抗酸杆菌(AFB)涂片显微镜检查技术质量,(2)分析操作过程误差对检查结果的影响.设计对抗酸杆菌(AFB)涂片显微镜检查的督导评价结果进行分析.评价标本、涂片、染色以及结果判读质量,并分析这些参数之间的联系.结果优良质量标本的比例比较高.研究结果证明,阳性结果与黏液脓性痰标本之间有直接联系.薄涂片的比例相对较高.薄涂片的阳性率以及细菌计数较低.染色质量良好.全国的读片一致性为98%.然而假阳性率也比较突出,46%的假阳性与染色缺陷有关.结论阿根廷结核病实验室网络的抗酸杆菌涂片显微镜检查技术质量以及涂片判读一致性比较满意.然而,需要在以下方面加以改进涂片质量、染色和读片、覆盖率、督导工作的非中心化、涂片的选择方法以及数据登记等.也需要开展涂片保存方面的实施性研究.     

1983-2001年阿根廷结核病实验室网络抗酸杆菌涂片显微镜检查的质量评价

地点:阿根廷结核病实验室网络.目标:(1)评价1983-2001年间督导的抗酸杆菌(AFB)涂片显微镜检查技术质量,(2)分析操作过程误差对检查结果的影响.设计:对抗酸杆菌(AFB)涂片显微镜检查的督导评价结果进行分析.评价标本、涂片、染色以及结果判读质量,并分析这些参数之间的联系.结果:优良质量标本的比例比较高.研究结果证明,阳性结果与黏液脓性痰标本之间有直接联系.薄涂片的比例相对较高.薄涂片的阳性率以及细菌计数较低.染色质量良好.全国的读片一致性为98%.然而假阳性率也比较突出,46%的假阳性与染色缺陷有关.结论:阿根廷结核病实验室网络的抗酸杆菌涂片显微镜检查技术质量以及涂片判读一致性比较满意.然而,需要在以下方面加以改进:涂片质量、染色和读片、覆盖率、督导工作的非中心化、涂片的选择方法以及数据登记等.也需要开展涂片保存方面的实施性研究.     

显微镜下多血管炎的诊断和治疗

显微镜下多血管炎（MPA）是一种损害小动脉、小静脉及毛细血管的少见疾病，男性多见，平均发病年龄在50岁以上。MPA可以累及多个脏器，尤其是肾脏，临床上主要表现为快速进展的肾小球肾炎等系统性损害。MPO－ANCA的阳性率高是其重要的免疫学特征，而免疫荧光检查血管壁无或仅有微量的免疫成分沉积。肾活检是MPA区别于其他血管炎的鉴别要点。MPA的治疗可联合使用肾上腺糖皮质激素和细胞毒药物。     

显微镜下多血管炎的临床和影像学诊断体会

目的探讨显微镜下多血管炎（microscopic polyangiitis,MPA）肺部表现的临床特点及误诊原因。方法对有肺部表现的3例MPA患者的诊断过程、误诊原因和胸部影像学表现进行回顾性分析。结果3例MPA患者都有肺部损害,以发热、咳嗽、咯血、呼吸困难为主要临床表现,胸部影像主要表现为两肺弥漫性斑片状或大片磨玻璃样和实变阴影。3例MPA患者抗中性粒细胞胞浆抗体阳性。确诊前曾被误诊为肺炎、肺结核、间质性肺炎等。确诊后给予甲基强的松龙联合环磷酰胺冲击治疗,临床症状及肺部表现近期内迅速好转。结论肺是MPA的常见累及脏器之一。误诊原因主要是对本病及其肺损害的特点认识不足。     

经电子支气管镜对14例肺及支气管结核的诊断

目的探讨支气管镜诊断肺及支气管结核的价值。方法对14例经电子支气管镜确诊的肺及支气管结核患者的临床表现、辅助检查和支气管镜结果进行分析。结果支气管镜结果显示，炎症浸润型6例，干酪样坏死型6例，肉芽增殖型2例，刷检涂片及术后痰找抗酸杆菌阳性12例。结论支气管镜应为支气管结核最重要也是最有效的方法。     

不同结核分支杆菌抗原对结核病的诊断价值

目的　研究不同结核分支杆菌单体蛋白抗原在结核病体液免疫诊断中的价值。方法以快速免疫色谱法（ＩＣＴＴＢ卡） 检测结核分支杆菌５ 种单体蛋白的抗原性，观察其在结核病诊断中的敏感性和特异性，并与ＰＰＤ 皮试及脂阿拉伯糖甘露糖（ＬＡＭ） 相比较。结果　ＩＣＴＴＢ 卡、ＬＡＭ 及ＰＰＤ皮试诊断结核病的敏感性分别为５５．９％ 、５２．６％ 、７５．０％ ，后者与前二者间差异均有显著意义（ Ｐ＜０ ．０５）；ＩＣＴＴＢ卡、ＬＡＭ、ＰＰＤ皮试诊断肺结核的特异性为８７ ．６％ 、６２．９％ 、６８．９％ ，前者与后二者间差异均有非常显著意义（ Ｐ＜ ０．０１）；ＩＣＴＴＢ卡诊断肺结核病的临床阳性预计值为８８ ．５％ ，临床阴性预计值为５３ ．８ ％ ；非结核病例中的ＩＣＴＴＢ阳性主要由抗原４、５ 阳性所致，如果去除抗原４ 、５，那么ＩＣＴＴＢ诊断结核病的特异性将提高到９８．９％ 。结论　血清结核分支杆菌单体蛋白抗原性的测定是肺结核病一项有用的辅助诊断手段。     

糖脂抗原和包膜抗原血清学检测活动性结核IgG抗体价值评估

目的评估脂阿拉伯甘露糖─ＩｇＧ（ＬＡＭ－ＩｇＧ）和包膜蛋白─ＩｇＧ对活动性结核的诊断价值。方法两种抗原分别采用快速和常规酶朕免疫吸附法（ＥＬＩＳＡ）进行检测。选择１０８例活动性结核（其中９７例活动性肺结核、１１例肺外结核），６９例非结核肺部疾患（其中肺癌１６例，其它肺部疾患５３例）血清平行检测结核分支杆菌ＩｇＧ抗体。结果９７例活动性肺结核涂阳敏感性分别为７１．８％（２８／３９）和９２．３％（３６／３９），涂阴为６０．３％（３５／５８）和７７．６％（４５／５８）。６９例非结核肺部疾患组两法阳性分别为４例和１４例，假阳性率分别为５．８％（４／６９）和２０．３％（１４／６９）；特异性分别为９４．２％（６５／６９）和７９．７％（５５／６９）。结论提示快速ＥＬＩＳＡ血清ＬＡＭ－ＩｇＧ测定操作简便、快速、不需任何仪器设备且特异性较高，对活动性结核病有较高的临床辅助诊断价值。     

结核分枝杆菌重组EIS蛋白血清学诊断价值的研究

目的研究结核分枝杆菌重组EIS蛋白的抗原性,观察抗结核化疗对抗EIS抗体滴度影响,初步探讨EIS蛋白作为血清诊断抗原在筛选活动性肺结核作用的可能性.方法采用基因工程方法表达纯化EIS抗原,以EIS为抗原建立酶联免疫方法,检测肺结核病人和健康对照者中抗EIS抗体的存在情况.对初次化疗的肺结核病人进行抗EIS抗体水平的随访,观察化疗对抗体反应的影响.结果肺结核病人抗EIS抗体的水平显著高于健康对照和肺部非肺结核疾病对照（P＜0.000 1）,结核化疗对抗体反应的影响不大,在整个化疗过程中抗体水平无显著差别.结论 EIS抗体的检测可能有助于活动性肺结核的筛选.     

Integration of microscopy and serodiagnostic tests to screen for active tuberculosis.

SETTING: University of California San Diego Medical Center, USA. OBJECTIVE: To create a simple screening strategy for tuberculosis (TB) that includes antibody detection assays to improve the accuracy of microscopic examination of sputum for acid-fast bacilli (AFB smear). METHODS: Serum samples were obtained from 190 patients suspected of having active TB. TB diagnosis was established by Mycobacterium tuberculosis culture. HIV status was determined by commercial serologic tests. IgG antibody levels were measured by ELISA using purified M. tuberculosis antigens. Data from 130 randomly selected patients were used to develop a screening strategy; data from the remaining 60 patients were used for validation. RESULTS: AFB smear had 70% sensitivity and 88% specificity. In algorithms integrating single or multi-antigen ELISA with AFB smear and HIV results, the sensitivity improved over each test alone. The algorithm that included a four-antigen ELISA (38 kDa antigen, lipoarabinomannan, MPT-64 and glutamine synthase) had a sensitivity of 93% and a specificity of 76%. Compared to AFB smear, the sensitivity of the algorithm was significantly higher, while the specificity was not statistically different. CONCLUSION: This study demonstrates that a screening strategy can be created by integrating multi-antigen ELISA with AFB smear and HIV testing.     

结核抗体检测对活动性肺结核的诊断价值评价

目的 评价结核抗体检测对活动性肺结核的诊断价值。方法 收集2016年8月至2017年8月在重庆市公共卫生医疗救治中心结核科住院的495例活动性肺结核患者(观察组)和158例非结核呼吸道疾病患者(对照组),均为综合患者的临床表现、胸部影像、痰细菌病原学检测或诊断性抗结核药物治疗有效等资料临床确诊后进行血清学诊断。分析血清MTB免疫球蛋白G(immunoglobulin G,IgG)、免疫球蛋白M(immunoglobulin M,IgM)、脂阿拉伯甘露聚糖(lipoarabinomannan,LAM),以及相对分子质量16000(以下采用“16kD”表示)和相对分子质量38000(以下采用“38kD”表示)的蛋白抗体的检测资料,以及单独及联合检测不同结核抗原(LAM、38kD和16kD)的结果,评价两组患者结核抗体检测的敏感度、特异度、阳性预测值、阴性预测值,以及对活动性肺结核的诊断效能。结果 495例观察组患者血清结核抗体检测阳性率[68.7%(340/495)]明显高于158例对照组患者[34.8%(55/158)],差异有统计学意义(χ ²=57.50,P<0.01);菌阴肺结核患者血清结核抗体检测阳性率[64.0%(210/328)]明显低于菌阳肺结核患者[77.8%(130/167)],差异有统计学意义(χ ²=9.83,P<0.01)。观察组340例结核抗体阳性患者中,LAM、38kD、IgG抗体联合检测同时均阳性的患者最多[61.8%(210/340)];对照组55例结核抗体阳性患者中,单一IgG抗体阳性最高[67.3%(37/55)]。以临床诊断为标准,血清结核抗体对活动性肺结核的诊断敏感度、特异度、阳性预测值和阴性预测值、总符合率、约登指数分别为68.7%(340/495)、65.2%(103/158)、86.1%(340/395)和39.9%(103/258)、67.8%(443/653)、0.34。 结论 血清结核抗体检测活动性肺结核患者具有较高的阳性率和敏感度,对诊断活动性肺结核具有一定的辅助价值,其中菌阳肺结核患者的检测阳性率高于菌阴肺结核,LAM、38kD和IgG联合检测可提高活动性肺结核的诊断阳性率。     

Evaluation of serodiagnostic tests for visceral larva migrans.

Four serologic techniques for the diagnosis of visceral larva migrans caused by Toxocara canis, namely indirect hemagglutination (IHA), bentonite flocculation (BF), enzyme-linked immunosorbent assay (ELISA), and double diffusion in agar (Ouchterlony), were evaluated using sera sent to the Center for Disease Control from patients with a presumptive diagnosis of visceral larva migrans (VLM). Patients having 5-6 of the clinical or laboratory criteria for VLM were designated as cases while those with 0-2 criteria served as controls. The sensitivity of the ELISA was 78.3% compared to 18.2%, 25.8% and 65.2% for the IHA, BF, and Ouchterlony, respectively; the sepcificity of all four tests was greater than 92%. The predivtive value of positive test was greater than 85% for all tests except the IHA, while the predictive value of a negative test was greater than 85% only for the ELISA. The results of a ELISA were reproducible in different laboratories. Based on these findings, the ELISA using a larval antigen appears to be the serodiagnostic method of choice for VLM.     

Role of serodiagnostic tests and skin tests in the diagnosis of fungal disease

Serodiagnostic tests have a significant role in the diagnosis of a number of fungal infections. The bases for these tests are reviewed and their sensitivity, specificity, and applications are discussed. Skin test antigens are also reviewed. Their diagnostic role is currently limited, but they have some utility as epidemiological tools.     

Side-room tests to screen for microalbuminuria in diabetes mellitus

Three side-room tests (latex bead immunoagglutination test, LBT; 25% sulphosalicylic acid test, SST; microalbutest, MAT) for the detection of microalbuminuria in diabetics are described and their screening potential and practicability assessed. One hundred insulin-dependent diabetics attending a diabetic clinic provided an early morning urine sample (Albustix-negative) which was subjected to each of the three tests, and urinary albumin concentration (UA) was assayed by RIA. Tests were assessed in random order by two trained operators using a semiquantitative grading scale with 100% concordance between 10 observers. All test results greater than or equal to trace +ve were sufficiently sensitive (sensitivity greater than or equal to 90%) in detecting UA greater than 15 mg/l, but MAT exhibited a significantly reduced specificity (69%) and positive predictive value (58%). For a reference UA greater than 30 mg/l, LBT and SST results greater than or equal to trace +ve and MAT results greater than or equal to +ve showed a sensitivity of 100%, a specificity greater than 85% and a positive predictive value greater than 60%. Reagent shelf-life was shortest with LBT. SST involved centrifugation or filtration. Technical skill required was highest with LBT and lowest with MAT. Costs were slightly higher with LBT than SST and were not available for MAT.     

CysA2: A candidate serodiagnostic marker for Mycobacterium tuberculosis infection

Background and objective: It is important to identify and test serologically active antigens in order to devise a mixture of antigens or peptides that is most useful for serodiagnosis. This study evaluated the serodiagnostic potential of CysA2, which has not previously been described as a serological antigen, together with those of PstS1, HspX, antigen 85 complex and CFP‐10 proteins. Methods: Serum IgG antibody titres against each antigen and a mixture of the antigens were measured by ELISA, in subjects with pulmonary tuberculosis and in healthy control subjects. Results: CysA2 showed diagnostic value comparable to that of PstS1 and HspX. Mixtures of these three proteins provided the highest diagnostic sensitivity. CysA2 was useful for identifying patients who did not react to HspX or PstS1, and was most valuable in increasing the sensitivity of testing. Furthermore, CysA2 efficiently overcame the limitation associated with use of PstS1, that is, significantly lower sensitivity for subjects who are negative for acid‐fast bacilli. Conclusions: These findings suggest that CysA2 can be used in combination with HspX and/or PstS1 to increase the accuracy of tuberculosis diagnoses.     

三种结核分枝杆菌分泌抗原的血清学诊断价值

目的 研究3种结核分枝杆菌分泌抗原（相对分子质量分别为81000和38000的抗原、CF-10抗原）在结核病血清学诊断中的价值。方法 收集安徽省2002年3月至2005年4月结核病患者的血清标本149例，其中痰涂片阳性结核病患者108例，痰涂片阴性结核病患者41例；非结核性肺部疾病患者血清标本24例；健康对照170例。取指数生长中晚期结核分枝杆菌培养液，纯化38000天然抗原，以和6个组氨酸融合的形式表达，镍离子螫合的层析柱亲和纯化81000抗原和C-10抗原，通过酶联免疫吸附测定方法检测3种结核分枝杆菌分泌抗原的抗原性与特异性。结果 纯化的抗原具备较高的纯度（〉95％）和检测特异性（〉98％）。结核病患者的81000抗原、38000抗原和CF-10抗原阳性检出率分别为64．4％（96／149）、57．0％（85／149）和37.6％（56／149），阴性符合率均大于98％。3种抗原联合使用对结核病患者的阳性检出率达到81．2％（121／149），阴性符合率大于98％。结论 3种抗原均具有较好的特异性，3种抗原联合使用具有良好的敏感性与特异性，明显优于任何一种抗原单独使用的效果，将多个抗原联合使用是进行高质量的结核分枝杆菌血清学诊断切实可行的手段。     

Bacteriological tests for tuberculosis

This review describes current developments for the bacteriological diagnosis of active tuberculosis. It deals mainly with molecular methods, describing their performance and how they can be integrated into more traditional diagnostic approaches. At present, microscopic examination and culture are still essential for the diagnosis of TB and to guide therapeutic decisions. Nucleic acid amplification and line probe assays speed up the identification and susceptibility testing of mycobacteria in AFB smear positive specimens or in culture. They are also efficient for comparison of M.?tuberculosis strains with each other (genotyping). On the other hand, at present, molecular tests are not applicable for diagnosis in smear negative specimens and even less so for diagnosis of culture-negative tuberculosis. The use of serology for antibody/antigen detection is not useful and it is not appropriate to assays based on the release of interferon-γ release as they are currently available. Notable progress has been made but more sensitive diagnostic tests for TB are still urgently needed.