半乳甘露聚糖抗原检测法对血液患者侵袭性真菌感染早期诊断的价值

目的评价半乳甘露聚糖抗原检测法对血液肿瘤患者侵袭性检测真菌感染早期诊断的价值。方法 60例血液肿瘤患者中临床诊断为侵袭性真菌感染及拟诊断为侵袭性真菌感染48例,排除12例。在系统性抗真菌治疗前采集血液标本,采用ELISA试剂盒检测患者血清GM,计算GM试验的灵敏度、特异度、阳性和阴性预测值,观察系统性抗真菌治疗后GM水平的变化。结果按照试剂盒推荐的分界值（≥0.50ng/ml）时,GM试验诊断的灵敏度为77.5%,特异性为93.9%,阳性预测值为86.7%,阴性预测值为83.6%,按此标准准确定为临床血液肿瘤患者侵袭性真菌感染患者23例,拟诊侵袭性真菌感染患者25例。结论 GM试验对于血液肿瘤患者侵袭性真菌感染早期诊断具有重要的意义,且GM浓度与患者疾病的预后具有相关性。     

半乳甘露聚糖抗原检测法对血液患者侵袭性真菌感染早期诊断的价值

目的 评价半乳甘露聚糖抗原检测法对血液肿瘤惠者侵袭性检测真菌感染早期诊断的价值.方法 60例血液肿瘤患者中临床诊断为侵袭性真菌感染及拟诊断为侵袭性真菌感染48例,排除12例.在系统性抗真菌治疗前采集血液标本,采用ELISA试剂盒检测患者血清GM,计算GM试验的灵敏度、特异度、阳性和阴性预测值,观察系统性抗真菌治疗后GM水平的变化.结果 按照试剂盒推荐的分界值(≥0.50ng/ml)时,GM试验诊断的灵敏度为77.5%,特异性为93.9%,阳性预测值为86.7%,阴性预测值为83.6%,按此标准准确定为临床血液肿瘤患者侵袭性真菌感染患者23例,拟诊侵袭性真菌感染患者25例.结论 GM试验对于血液肿瘤患者侵袭性真菌感染早期诊断具有重要的意义,且GM浓度与患者疾病的预后具有相关性.     

铜绿假单胞菌胶体金免疫层析法的研制和应用

为建立一种简单、快速、准确的铜绿假单胞菌检测方法,结合抗重组铜绿假单胞菌外膜蛋白单克隆抗体（mAb）、胶体金标记技术,采用双抗体夹心法,建立了检测铜绿假单胞菌的胶体金免疫层析法,并对该方法的特异性、灵敏度和稳定性进行了评价,对临床痰标本进行了检测。结果显示,该胶体金免疫层析法能在3～15min内完成检测,方法的特异性高、稳定性强,检测灵敏度能达到105CFU/mL。与传统细菌培养方法相比,胶体金免疫层析法的相对灵敏度和特异性分别是80%和90.6%,两种方法的总符合率为86%。结论：利用抗铜绿假单胞菌单克隆抗体建立的胶体金免疫层析法,适合现场快速、特异检测铜绿假单胞菌。     

血清HWP1检测在儿童皮肤念珠菌病诊断中应用研究

目的 分析讨论血清白色念珠菌菌丝壁蛋白1 (hyphal wall protein 1,HWP1)检测在儿童皮肤念珠菌病诊断中的灵敏度和特异性等诊断效力.方法 选取本院2014年1月至2016年12月期间收治的86例儿童皮肤病患者进行研究,所有患儿均接受血清HWP1检测,将检测结果与金标准诊断结果对比,分析血清HWP1检测诊断儿童皮肤病念珠菌感染的灵敏度、特异性、阳性诊断价值和阴性诊断价值.结果 本研究中86例患几经组织培养确诊为念珠菌感染的患儿41例,非念珠菌感染患儿45例,两性间血清HWP1抗体阳性率相比,差异无统计学意义(P＞0.05);念珠菌感染患儿血清HWP1抗体阳性率(85.37％)显著高于非念珠菌感染患儿(6.67％),差异有统计学意义(P＜0.05);血清HWP1检测诊断灵敏度85.37％,特异性93.33％,阳性预测值92.11％,阴性预测值87.50％,ROC曲线下面积为0.896.结论 血清HWP1能够为儿童皮肤病念珠菌感染筛查和鉴别诊断提供高价值信息,其灵敏度优良,特异性非常高,值得临床推荐.     

血浆(1-3)-β-D-葡聚糖检测对侵袭性肺部真菌感染的诊断价值

为评价(1-3)-β-D-葡聚糖检测(G试验)对侵袭性肺部真菌感染的诊断价值,对肺部感染患者130例行G试验,并将试验结果与病原学诊断结果相比较,计算G试验的灵敏度、特异性、阳性预测值、阴性预测值及准确性.结果表明:G试验对侵袭性肺部真菌感染的灵敏度为76.5%(13/17),特异性为80.7%(88/109),阳性预...     

1, 3-β-D葡聚糖、甘露聚糖IgM/IgG抗体对侵袭性念珠菌病早期诊断的临床价值

目的评估1, 3-β-D葡聚糖(BDG)、甘露聚糖IgM抗体(Mn-IgM)、甘露聚糖IgG抗体(Mn-IgG)对侵袭性念珠菌病的诊断价值, 对比分析血清学标志物独立及联合检测在诊断性能上的差异。方法收集126例侵袭性念珠菌病患者和104例同期健康体检者的血清作为研究对象。采用动态显色法检测BDG, ELISA法检测Mn-IgM、Mn-IgG。比较分析3项血清学标志物独立及联合检测的敏感性、特异性、阳性预测值、阴性预测值、约登指数、符合率、Kappa值, 绘制受试者工作特征(ROC)曲线并计算ROC曲线下面积(AUC)。结果 BDG、Mn-IgM、Mn-IgG在侵袭性念珠菌病患者中明显高于健康体检人群(P<0.01)。BDG的敏感性和特异性分别为48.41%、92.31%, Kappa值为0.389, AUC为0.842;Mn-IgM的敏感性和特异性分别为64.29%、91.35%, Kappa值为0.540, AUC为0.829;Mn-IgG的敏感性和特异性分别为27.78%、95.19%, Kappa值为0.214, AUC为0.737。BDG+Mn-IgM的敏感性和特异性分...     

重组汉坦病毒核衣壳蛋白抗原用于胶体金标记免疫层析法检测肾综合征出血热抗体

目的制备高表达量、高活性的HV NP重组抗原,以此为基础建立一个可同时检测HFRs患者血清中IgM和lgG抗体的快速胶体金标记诊断试剂盒。方法从TA-A537中扩增出截短的HV S基因片段P6-119,定向克隆至原核表达载体pET-30a中进行原核表达,采用亲和层析法纯化重组蛋白。以胶体金标记重组抗原,应用金标快速免疫层析法同时检测HFRS患者血清中IgM和IgG抗体。结果金标快速免疫层析法可同时检测HFRS患者血清中IgM和IgG抗体,与IFA比较,IgG抗体检测的敏感性和特异性分别为94.9%、100%。与ELISA比较,IgM抗体检测的敏感性和特异性均为100%。结论截短的重组汉坦病毒NP蛋白表达量高且具有良好的抗原性。以此为基础建立的金标快速免疫层析法显示出很高的敏感性和特异性,且具有简便、快速等优点,非常适用于各种层次尤其是缺乏实验条件和专业人员的基层医疗单位对HFRS疑似患者作出早期诊断。     

侵袭性念珠菌病实验室诊断现状的分析与思考

侵袭性念珠菌病是第四位医院血源感染性疾病,迄今缺乏理想的诊断方法。尽管以念珠菌抗原、抗体和核酸为靶标的检测方法的研究取得了很大的进展,但由于敏感性、特异性、标准化以及实验设计等原因,难以解决临床实际问题。高度保守、含量丰富的诊断标志物可能是解决这一问题的良好出路。     

霍乱弧菌O1胶体金免疫层析快速检测法的建立

目的建立一种快速、简易的检测霍乱弧菌O1群的胶体金免疫层析法(GICA)。方法利用胶体金免疫层析技术,采用双抗体夹心法检测霍乱弧菌O1群,对该法进行敏感性、特异性和稳定性分析。与细菌培养法对比检测208份临床标本。结果该法能在10分钟内完成检测;该试纸条仅与霍乱弧菌O1群阳性样品发生特异性反应;检测霍乱弧菌O1群的最低检出浓度为1×105cfu/mL;与细菌培养法对比检测208份临床标本,特异性和灵敏度均达100%。结论新建立的霍乱弧菌O1群胶体金免疫层析试验简便、快速,特异性和灵敏度较好,适用于现场样品的快速筛查。     

念珠菌耐药机制的研究进展

近年来,随着侵袭性真菌感染危险因素的增多,如器官移植、中性粒细胞减少、长期应用糖皮质类固醇激素等,侵袭性真菌感染的发病率和死亡率不断上升。临床上侵袭性真菌感染仍以念珠菌属感染为主,其中白念珠菌感染占54％、光滑念珠菌占22％、近平滑念珠菌占6％、热带念珠菌占5．7％、克柔念珠菌占4．3％。随着抗真菌药物长期反复广泛应用,对抗真菌药物耐药的念珠菌也愈来愈多,     

Diagnosis of invasive candidiasis by a dot immunobinding assay for Candida antigen detection.

A dot immunobinding assay which uses a polyclonal rabbit anti-Candida immunoglobulin G as the primary antibody and colloidal gold coated with goat anti-rabbit immunoglobulin G as the secondary antibody for the detection of Candida cytoplasmic antigens is described. It was able to detect as little as 1 ng of total Candida protein per ml when a cytoplasmic extract of Candida albicans was seeded into buffer and 10 ng/ml when the same extract was seeded into pooled human serum. Serial serum samples from four groups of patients were assayed for Candida antigen: (i) 22 patients with candidemia, (ii) 16 patients at high risk for invasive candidiasis, (iii) 3 patients with other deep mycoses, and (iv) 50 hospitalized patients at low risk for serious Candida infection. Of the 22 candidemic patients, 19 had invasive candidiasis and 3 had transient candidemia. Antigenemia was detected in 16 of the 19 patients with invasive candidiasis (including patients with C. albicans, Candida tropicalis, Candida glabrata, Candida krusei, and Candida parapsilosis) and in 4 of 16 patients at high risk for invasive candidiasis. There was no detectable antigen in 12 high-risk control patients, 3 patients with transient candidemia, 3 patients with other deep mycoses, and 50 relatively low-risk patients. The sensitivity for detecting invasive disease in candidemic patients and specificity for all patients studied were 84.2 and 94.4%, respectively. The positive predictive value was 80%; the negative predictive value was 95.7%. The sensitivity for neutropenic patients with invasive disease was 85.7%. This assay is rapid and accurate and appears to be useful in identifying candidemic patients with invasive candidiasis.     

Assessment of analysis of urinary pneumococcal antigen by immunochromatography for etiologic diagnosis of community-acquired pneumonia in adults

The limitations of conventional microbiologic methods (CMM) for etiologic diagnosis of community pneumococcal pneumonia have made faster diagnostic techniques necessary. Our aim was to evaluate the usefulness of the immunochromatography (ICT) technique for detecting urinary Streptococcus pneumoniae antigen in the etiologic diagnosis of community-acquired pneumonias (CAP). This was a prospective study on in-patients with CAP in a tertiary hospital conducted from October 2000 to March 2004. Apart from using CMM to reach an etiologic diagnosis, we determined pneumococcal antigen in concentrated urine by ICT. We also determined the urinary pneumococcal antigen (UPA) content in patients from two control groups to calculate the specificity of the technique. One group was comprised of in-patients diagnosed with chronic obstructive pulmonary disease (COPD) or asthma, with respiratory infection, and without pneumonia; the other group included fractures. We studied 959 pneumonia patients and determined UPA content in 911 (95%) of them. We diagnosed the etiology of 253 cases (28%) using CMM; S. pneumoniae was the most common etiologic agent (57 cases). ICT analysis was positive for 279 patients (31%). Using this technique, the percentage of diagnoses of pneumococcal pneumonias increased by 26%, while the overall etiologic diagnosis increased from 28 to 49%. The technique sensitivity was 81%; the specificity oscillated between 80% in CAP with nonpneumococcal etiology and 99% for patients with fractures without infections. Determination of UPA is a rapid, simple analysis with good sensitivity and specificity, which increased the percentage of etiologic diagnoses. Positive UPA may persist in COPD patients with probable pneumococcal colonization or recent pneumococcal infections.     

Diagnosis of Streptococcus pneumoniae pneumonia by quantitative enzyme linked immunosorbent assay of C-polysaccharide antigen.

AIMS--To evaluate the use of a quantitative enzyme linked immunosorbent assay (ELISA) detecting C-polysaccharide (PnC) antigen in sputum for the diagnosis of Streptococcus pneumoniae infection. METHODS--Specimens of sputum from 60 patients with acute community and hospital acquired pneumonia and infective exacerbations of obstructive airways disease were examined by semiquantitative culture and antigen ELISA. RESULTS--Using a cutoff value of 1 microgram/ml PnC antigen for a positive result, the sensitivity of this assay was 90.3%, specificity 93.1%, predictive value of a positive result was 93.5%, and the predictive value of a negative result 89.6%. CONCLUSIONS--Quantitation of C-polysaccharide antigen in sputum by ELISA distinguishes between carriage of oral bacteria which express PnC-like antigen and infection with S pneumoniae and compares favourably with other diagnostic methods.     

Invasive pulmonary aspergillosis complicating chronic obstructive pulmonary disease in an immunocompetent patient

Immunocompromised individuals are susceptible to pulmonary aspergillus infection, but invasive aspergillus infection is extremely rare in the presence of normal immunity. We report a case of invasive aspergillosis in an immunocompetent 63-year-old male with chronic obstructive pulmonary disease (COPD). Patients with COPD may be at risk for developing pulmonary aspergillus infection, which should be considered as a diagnostic possibility in patients with unresolving pulmonary infection.     

Pulmonary aspergillosis in an unselected autopsy series

The present study was performed to analyze the relationship between underlying diseases and the morphologic form of aspergillosis. This retrospective analysis of 3284 autopsies yielded 18 cases of aspergillosis. The specific diagnosis of aspergillosis was rendered by a monoclonal antibody versus Aspergillus spp. Patients with hematological disorders, such as acute leukemia and aplastic anemia, made up about 35% of all patients dying of invasive aspergillosis. Diseases of the airways and the pulmonary parenchyma constituted the second most pathogenetic factor for the development of aspergillosis. The morphologic form of aspergillosis was closely related to the underlying diseases. Non- and semi-invasive forms of aspergillosis--saprophytic infection and chronic necrotizing aspergillosis--were observed only in patients with an isolated underlying pulmonary disease devoid of any other precipitating factor. In contrast, seven patients, five of whom suffered from hematological diseases, had no underlying lung disease and developed aspergillus pneumonia. The remaining 5 patients with aspergillus pneumonia showed a combination of underlying extrapulmonary disease and pulmonary alterations that preceded aspergillosis. The local distribution of fungal infection showed a characteristic distribution pattern with a predominance of the upper lung lobes. Hematogeneous spread beyond the lungs occurred exclusively in cases with aspergillus pneumonia. We conclude that the different forms of aspergillosis are closely related to the nature of the underlying disease.     

斑点金免疫渗滤法检测血清抗精子抗体

目的建立检测血清抗精子抗体的斑点金免疫渗滤法 (dot im munogold filtration assay,DIGFA)。方法　采用经 Sephadex G- 75 -抗人全血清柱亲和层析纯化的人精子抗原和胶体金标记的 SPA,根据免疫渗滤原理 ,建立抗精子抗体(ASAb)的斑点金免疫渗滤法检测法。结果　纯化的人精子抗原具有较好的特异性 ;斑点金免疫渗滤法与 EL ISA法比较 ,符合率为 97.7%。特异性为 96 .2 % (TN/ TN+ FP× 10 0 % ) ,敏感性为 88.9% (TP/ TP+ FN× 10 0 % ) ,阳性预测值 84.2 % (TP/TP+ FP) ,阴性预测值 97.2 % (TN/ TN+ FN) .结论　斑点金免疫渗滤法方法检测血清抗精子抗体有较好的特异性和敏感性 ,重复性好 ,试剂稳定 ,操作简便快速 ,具有临床应用价值     

免疫层析法同时检测孕妇TORCH—IgM抗体的研究

目的建立一种免疫层析法用于同时检测孕妇血清中TORCH—IgM抗体。方法用胶体金标记羊抗人IgM（斗链）抗体，将巨细胞病毒（CMV）、风疹病毒（RV）、弓形体（Tox）、单纯疱疹病毒Ⅱ型（HSV-Ⅱ）四种重组抗原同时包被在硝酸纤维膜上，研制出同时检测TORCH—IgM抗体免疫层析试纸条。结果自制的免疫层析法与ELISA法对比检测阴阳性血清标本，各单项指标抗-CMV—IgM、抗-RV—IgM、抗-Tox—IgM、抗-HSV-Ⅱ-IgM阳性符合率分别为85．7％、100％、100％、100％，两法总符合率分别为97．9％、100％、100％、100％。结论本方法操作简便，显示结果直观，并且特异、稳定、快速、重复性好，可用于孕妇优生优育检测。     

早期诊断心肌损伤肌球蛋白胶体金免疫层析法研究

目的建立一种简便、快速、准确检测人心肌损伤的胶体金免疫层析法（GICA）。方法制备胶体金标记抗Myosin多肽抗体，抗Myosin单克隆抗体，结合垫和样品垫的处理，组装免疫层析试纸条。检测患者血清中Myosin，进行敏感性和特异性评定。结果测试条灵敏度可达5ng／mL。检测30例急性心肌梗塞（AMI）患者血清Myosin水平，并与Roche肌钙蛋白胶体金免疫层析试剂条比较，符合率达84.2％。结论本方法特异性强、灵敏度高、简便快速、可用于急性心肌损伤的早期诊断。