国产血清半乳甘露聚糖检测试剂对侵袭性肺曲霉菌病的诊断价值评估

目的：评估国产血清半乳甘露聚糖（Galactomannan,GM）检测试剂对侵袭性肺曲霉菌病的诊断价值。方法根据血液病/恶性肿瘤患者侵袭性真菌病的诊断标准与治疗原则（第四次修订版）[1]收集临床确诊侵袭性肺曲霉菌病（inva-sive pulmonary aspergillosis,IPA）、临床诊断IPA、拟诊IPA、排除IPA四组病例。采用天津贻诺琦公司酶联免疫吸附法（ELISA）试剂检测纳入的86例患者血清标本的GM浓度,分析其敏感性、特异性、阳性预测值（PPV）、阴性预测值（NPV）。结果86例病例中,临床诊断27例、拟诊12例、排除47例。在3种不同的阳性判断标准下,敏感性：9444%、9630%、6296%;特异性：5625%、4576%、6441%;PPV：4474%、4483%、4474%;NPV：9643%、9643%、7917%。统计学分析证实标准1（即血清GM值〉095μg/L为阳性,〈075μg/L为阴性,075~095μg/L为灰区,未将灰区加入计算）在3种判断标准中最优,故选择其为最终判断标准。结论该血清GM检测试剂盒诊断性能较好,可以用于侵袭性肺曲霉菌病的辅助诊断。     

半乳甘露聚糖对侵袭性曲霉菌感染的诊断价值

近年来侵袭性曲霉菌感染（invasive aspergillosis,IA）发病率逐渐增多,已经成为免疫功能低下患者致死率高的主要原因。有报道称在造血干细胞移植（HSCT）患者中IA的发病率为2％～26％,     

侵袭性肺曲霉菌患者血清半乳甘露聚糖动态变化及与CT影像学关系

目的观察侵袭性肺曲霉菌感染患者抗真菌治疗前后肺泡灌洗液及血清中半乳甘露聚糖(GM)的水平变化、CT影像学特征及血清中性粒细胞数与GM试验阳性率之间的关系,患者血清中性粒细胞值与GM试验阳性率之间的关系。方法连续性纳入60例确诊侵袭性肺曲霉菌感染患者,观察抗真菌治疗前后肺泡灌洗液和血清GM试验动态水平变化。单因素Logistic回归分析CT侵犯肺叶数、空气新月征、晕轮征、血清中性粒细胞值与GM的关系。结果抗真菌治疗前患者肺泡灌洗液GM平均值均高于血清GM平均值(0.62±0.08mg/L,60.0%;0.51±0.07mg/L,31.6%)。同抗真菌治疗前比,患者抗真菌治疗后1个月、2个月、3个月、6个月肺泡灌洗液GM值快速降低(P0.05),而血清GM数值缓慢降低,仅在第3个月、第6个月降低(P0.05)。患者CT侵犯肺叶数、晕轮征与肺泡灌洗液、血清中性粒细胞值GM试验阳性率显著相关。结论肺泡灌洗液及血清GM试验可作为IFD疾病诊断及治疗的重要辅助指标。CT影像学特征、血清中性粒细胞值与GM试验阳性率显著相关。     

半乳甘露聚糖用于侵袭性曲霉感染实验诊断的研究现状

曲霉是导致深部真菌感染的重要条件致病菌,临床早期诊断和治疗对预后具有明确意义。ELISA检测血清中半乳甘露聚糖用于诊断曲霉感染具有良好的敏感度(64.5%～76%)和特异度(81%～98.7%),可用于曲霉感染的早期诊断及治疗的监测。     

血清半乳甘露聚糖试验在诊断马尔尼菲青霉病中的应用

目的评价血清半乳甘露聚糖(GM)试验在诊断马尔尼菲青霉感染(PSM)中的应用价值。方法收集2012~2015年间确诊PSM患者37例、其他真菌感染患者17例、健康志愿者30例血清,进行GM试验检测;同时评价其中4例PSM患者治疗前后GM值变化,计算诊断指标并分析评价。结果 GM试验对PSM患者总体敏感度为78.38%,特异度为82.98%;其中对HIV阳性PSM患者敏感度为76.47%,特异度为97.87%;对HIV阴性PSM患者敏感度为75.00%,特异度为84.78%。4例随访的PSM患者经抗真菌药物治疗后,随着病情好转,GM水平逐渐下降。结论 GM试验对PSM有一定的诊断价值,尤其对马尔尼菲青霉流行地区的HIV感染者,早期诊断PSM有重要意义。     

支气管肺泡灌洗液半乳甘露聚糖检测对侵袭性肺曲霉病诊断价值的meta分析

目的通过系统评价分析支气管肺泡灌洗液(BALF)半乳甘露聚糖(GM)检测对侵袭性肺曲霉病(IPA)的诊断准确性并确定最佳诊断界值。方法检索Pubmed、EMBASE、OVID、Cochrane Library、中国知网(CNKI)、万方、中国生物医学(CBM)等数据库,自建库至2018年10月31日的相关数据,不限制语言,纳入符合标准的26个研究(71组数据),对其进行质量评价、异质性分析,然后分析计算每个诊断界值的合并指标,绘制受试者工作特征曲线,最终确定最佳诊断界值。结果 BALF-GM(诊断界值为0.5时)诊断IPA(确诊和临床诊断IPA)的合并灵敏度为0.87(95%CI 0.84～0.89),合并特异度为0.79(95%CI 0.77～0.81),合并阳性似然比为4.60(95%CI 3.49～6.05),合并阴性似然比为0.19(95%CI 0.14～0.25),合并诊断比值比为38.84(95%CI 22.99～52.81),集成受试者工作特征(SROC)曲线下面积(AUC)为0.92(95%CI 0.90～0.95)。在BALF-GM诊断界值为1.0时,受试者工作特征(ROC)曲线下面积(AUC)最大为0.94(95%CI 0.93～0.97)。结论 BALF-GM易于检测,对IPA的诊断准确性高,可以对IPA进行早期诊断,早期治疗,降低病死率,目前建议BALF-GM最佳诊断界值为1.0。     

半乳甘露聚糖检测在圈养海洋动物曲霉病早期诊断的应用

<正>曲霉菌是广泛存在于自然界的一种腐生菌。由于曲霉菌孢子较小,在空气中漂浮,一般是通过呼吸道进入机体感染后引起曲霉病,为条件性致病菌(刘正印和王爱夏,2007)。曲霉病在人类、家畜和许多野生物种中都有发现,引起从局部感染到致命以及对吸入孢子的过敏反应。曲霉病会引起海扇珊瑚致命性感染、蜜蜂僵化、鸟类肺部和气囊感染、牛真菌流产和乳腺感染、     

(1,3)-β-D-葡聚糖检测和半乳甘露聚糖抗原检测在侵袭性真菌病诊断中的价值探讨

目的 探讨(1,3)-β-D-葡聚糖检测和半乳甘露聚糖抗原检测在侵袭性真菌病诊断中的价值.方法 回顾性研究北京大学第一医院2008年1月～2010年7月临床疑诊侵袭性真菌病的住院患者.根据诊断标准确定是否诊断侵袭性真菌病.分析非培养诊断方法血清半乳甘露聚糖(GM试验)和血浆(1,3)-β-D-葡聚糖(G试验)的敏感度和特异度,以及二者联合应用后诊断的敏感度和特异度.结果 GM试验灵敏度为70％,特异度为84％;G试验灵敏度为50％,特异度为92％.GM试验和G试验二者联合试验时,其灵敏度和特异度分别为93％和78％.结论 GM试验和G试验均对侵袭性真菌病具有诊断价值,二者联合应用使其应用价值进一步提高.     

曲霉菌半乳甘露聚糖单克隆抗体的制备及初步应用

目的:制备抗曲霉菌半乳甘露聚糖的单克隆抗体,并基于获得的抗体建立用于快速准确检测曲霉菌感染的双抗体夹心酶联免疫吸附法(ELISA),以期可用于侵袭性曲霉菌病的临床诊断。方法:提取曲霉菌半乳甘露聚糖后免疫BALB/c小鼠,筛选与制备抗曲霉菌半乳甘露聚糖的单克隆抗体,通过间接ELISA法与Western Blot方法开展单克隆抗体检测性能分析,使用获得的单克隆抗体建立双抗体夹心ELISA方法,并初步应用于曲霉菌感染血清检测。结果:获得抗曲霉菌半乳甘露聚糖单克隆抗体5株,均可特异性识别曲霉菌半乳甘露聚糖,以其中性能最佳的3C9抗体和辣根过氧化物酶标记的3C9抗体配对为基础,建立了双抗体夹心ELISA方法,通过初步评价确定该方法可应用于临床侵袭性曲霉菌病血清检测,并且该方法与现有商品化试剂盒相比检测背景值较低,可更有效区分曲霉菌感染阴阳性血清。结论:本研究筛选获得针对曲霉菌半乳甘露聚糖的特异性单克隆抗体,以该抗体为基础建立双抗体夹心ELISA方法具有潜在转化应用前景,可为侵袭性曲霉菌病的临床诊断提供支持。     

内参标化的肺泡灌洗液GM试验对侵袭性肺曲霉菌病诊断效能的探讨

目的探讨肺泡灌洗液(BALF)半乳甘露聚糖检测(GM试验)内标物质的选择及内标化的方法,以及内标化后的BALF GM试验是否能够提高对侵袭性肺曲霉病(IPA)的临床诊断效能。方法收集2018年5月至2019年1月重庆医科大学附属第一医院临床拟诊为IPA住院患者的BALF标本156例,同时对曲霉菌抗原、钾离子(K~+)、尿素(UREA)、白蛋白(ALB)进行检测,依据组织病理、微生物结果、临床症状及特征、宿主因素等资料将研究对象分为IPA组39例和非IPA组117例,统计分析标化校正和未标化校正的BALF GM试验结果对IPA临床诊断效能的影响。结果标化校正和未标化校正的BALF GM试验I值在两组间均存在显著性差异(P0.001),经ROC曲线分析显示非标化BALF GM试验I值的cut-off值为0.69时,曲线下面积为0.731(P0.001),灵敏度71.8%,特异度70.1%;用K~+、UREA、ALB标化BALF GM试验I值后的ROC曲线分析,各cut-off值分别为0.605、1.905、0.69,曲线下面积分别为0.71、0.653、0.701 (P值分别为0.001、0.004、0.002),灵敏度分别为74.4%、66.7%、69.2%,特异度分别为70.9%、63.2%、69.8%。结论非标化BALF GM试验I值cut-off值为0.69时对IPA的诊断效能最高;用K~+标化后的BALF GM试验I值诊断IPA的效能优于非标化的BALF GM试验结果(P0.001)。     

Usefulness of galactomannan detection in the diagnosis and follow-up of hematological patients with invasive aspergillosis

The usefulness of galactomannan detection using the Platelia Aspergillus test for the diagnosis of invasive aspergillosis was studied in 849 sera from 54 hematological patients with prolonged neutropenia, which were classified according to the risk for invasive aspergillosis. Three patients developed a proven invasive aspergillosis, one a probable invasive aspergillosis and 17 patients a possible invasive aspergillosis. Thirty-three patients showed no evidence of invasive aspergillosis. All patients with proven invasive aspergillosis had a high risk for invasive aspergillosis, while the one having probable invasive aspergillosis had intermediate risk. Detection of galactomannan in this study showed a sensitivity of 66.7% for patients with proven invasive aspergillosis and 50% for patients with proven and probable invasive aspergillosis. The specificity was 98% or higher in all groups studied. The predictive positive and negative values for patients with proven invasive aspergillosis were 66.7% and 98%, respectively. A rise in the concentration of galactomannan was observed in patients who failed to respond to the antifungal treatment. Galactomannan antigenemia preceded post-mortem histological diagnosis of invasive aspergillosis in two patients by 17 and 81 days, respectively. In conclusion, detection of galactomannan by the Platelia Aspergillus test allows for a specific and relatively sensitive diagnosis of invasive aspergillosis in hematological patients with a high and intermediate risk for invasive aspergillosis.     

128例侵袭性肺曲霉病临床疗效分析

目的 通过回顾性病例分析,加强对侵袭性肺曲霉病的认识,重视对侵袭性肺曲霉病的分级诊断,从而制定合理治疗方案.方法 对四川大学华西医院201 1年1 ～12月128例诊断为侵袭性肺曲霉病患者的临床资料进行回顾性分析,重点分析基础疾病、临床表现、影像学表现、诊断及治疗方法.结果 确诊17例;依据宿主危险因素、临床表现及反复痰培养阳性拟诊111例.128例患者均伴有基础疾病.治疗方案包括伏立康唑（86例）,两性霉素B脂质体（1例）,伊曲康唑（12例）,米卡芬净（3例）,卡泊芬净（3例）,两性霉素B去氧胆酸盐（10例）,伏立康唑联合米卡芬净（1例）.治愈或好转81例（63.28％）,自动出院24例（18.75％）,死亡23例（17.97％）.结论 侵袭性肺曲霉病患者多数伴有多种感染危险因素,应当重视侵袭性肺曲霉病的分级诊断,据不同情况给予适当抗真菌治疗,以降低患者病死率.     

Early diagnosis of invasive aspergillosis in neutropenic patients with bi-weekly serial screening of circulating galactomannan by Platelia Aspergillus

The diagnosis of invasive aspergillosis in neutropenic individuals is difficult and lengthy since non-invasive diagnostic tests lack sensitivity and specificity. The diagnosis of invasive aspergillosis in 154 prolonged neutropenic patients was prospectively bi-weekly validated by screening circulating galactomannan. The global sensitivity was 73% and specificity was 96%. The positive and negative predictive values were 73% and 98% respectively. False positive reactions occurred at a rate of 2%. Antigenemia was detected before clinical suspicion of invasive aspergillosis (median, 6 days before) in 30% of patients and anticipated the onset of radiologic signs 9 days in 60% of patients. CONCLUSION: the prospective screening of galactomannan is a sensitive and non-invasive tool for early diagnosis of invasive aspergillosis in high-risk adult hematology patients.     

Value of Aspergillus galactomannan antigen detection in the diagnosis and follow-up of invasive aspergillosis in hematological patients

Serum galactomannan detection is considered to be a useful test for early diagnosis and follow-up of invasive aspergillosis. From February to September 2002, adult patients hospitalized in our Hematology Unit for receiving intensive chemotherapy and/or hematopoietic stem cell transplant were prospectively studied. We analyzed a total of 760 samples obtained from 100 patients. Eleven patients (11%) having a positive result (OD index >1.5 ng/ml) in two consecutive Platelia Aspergillus tests were considered galactomannan-positive cases. On the other hand, 12 patients (12%) were diagnosed of proven or probable invasive aspergillosis. Sensitivity (66.6%), specificity (95.5%), positive predictive value (72.7%) and negative predictive value (96.7%) were comparable to those of larger series. Galactomannan positivity allowed also to anticipate invasive aspergillosis diagnosis (from two to 17 days before radiographic findings and from two to 15 days before mycological culture). Moreover, kinetics of antigenemia could be useful for assessing therapeutic response. Once accepted galactomannan test as a diagnostic criterium for invasive aspergillosis knowing potential causes of false positive results is of paramount importance.     

Update on the contribution of galactomannan for the diagnosis of invasive aspergillosis

The diagnosis of invasive fungal infections (IFI) remains a challenge, particularly for diseases caused by filamentous fungi such as Aspergillus species. Unfortunately, many patients affected by these conditions are not identified before autopsy. Therefore, there is a need for new diagnostic methods for IFI. Galactomannan is a soluble antigen released during hyphal growth in tissues. A commercially available sandwich ELISA assay that detects galactomannan has been used in Europe for many years and is now approved for use in the USA. The test has an excellent negative predictive value in the detection of invasive aspergillosis (IA) in high-risk patients. In addition, it is more sensitive than culture and allows IA to be diagnosed before clinical manifestations occur. However, false-negative and false-positive results in certain populations are the main limitations to its use. The purpose of this review is to summarize the current knowledge about galactomannan testing in patients at risk for IA.     

The contribution of galactomannan detection in the diagnosis of invasive aspergillosis in bone marrow transplant recipients

Until recently, accurate microbiological diagnosis of invasive aspergillosis (IA) was seldom established in HSCT recipients. Blood samples are rarely positive for Aspergillus species, the reliability of the cultures depends of the specimen (if taken from a normally sterile site or not) and biopsy samples require invasive procedures, rarely recommended in patients with severe thrombocytopenia. Implementing the international consensus defining the microbiological criteria for the diagnosis of Aspergillus infection, we retrospectively evaluated the role of serum galactomannan (GM) detection by EIA to diagnose IA among HSCT patients with proven invasive fungal infection (IFI) and the impact of serum storage in GM concentrations. The EIA assay allowed categorizing as “probable” 5 of the 10 cases of “possible” aspergillosis (50%). Considering a lower cut-off level for the reaction (1.0), 80% of the cases could be categorized as “probable” aspergillosis. Positive or undetermined results were detected one to 4 months before the diagnosis of IA in eight of the 11 patients (72.7%) with proven IFI. Retesting the stored samples after a second storage for four years, we could observe lower reactivity in 20% of the samples. The detection of galactomannan by the EIA test represents a major advance in the diagnosis of IA in HSCT recipients at high risk of IA. A better understanding of the kinetics of the GM in different clinical situations is necessary to maximize the benefit of the test in Aspergillus surveillance.     

Evaluation of the diagnostic accuracy of galactomannan from the bronchoalveolar lavage fluid of patients with suspected invasive pulmonary aspergillosis

BackgroundSeveral studies to evaluate the accuracy of galactomannan (GM) in bronchoalveolar lavage fluid (BALF) as a diagnostic tool have been carried out; however, there are still controversies about the optimal cut-off point of BALF GM.AimsThe objective of this study was to determine the diagnostic accuracy and the optimal cut-off point on BALF GM from patients with suspected invasive pulmonary aspergillosis (IPA) in a tertiary care hospital.MethodsA cross-sectional study with 188 patients (≥18 years) that had undergone a bronchoscopy with BAL due to suspected IPA was carried out. IPA was diagnosed according to the EORTC/MSG guidelines.ResultsThe optimal optical density cut-off point for BALF GM was 0.67, with sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 70%, 32.3%, and 100%, respectively.ConclusionsBALF GM detection proved to be a useful supplementary technique in the early diagnosis of IPA in both neutropenic and non-neutropenic patients.     

肺部侵袭性曲霉菌感染的诊断思路

侵袭性真菌感染(invasive fungal infection,IFI)是免疫低下患者常见的并发症之一,尤其在血液肿瘤患者和造血干细胞移植后患者中,IFI更是影响患者生存和生活质量的重要因素。随着氟康唑在高危人群中的广泛预防性应用,白念珠菌的感染较前已有所减少,而曲霉菌的感染比例则有所增[1]