侵袭性真菌病实验室诊断方法临床应用专家共识

侵袭性真菌病的诊断引起临床多学科关注,早期确切诊断是改善患者预后的关键。其方法主要包括真菌直接镜检、真菌培养和鉴定、真菌血清学检查、分子生物学检测和组织病理学检查。为了促进临床医师深入了解并合理应用这些方法,提高侵袭性真菌病诊断水平,我们邀请多学科专家共同编写了此共识。本共识对侵袭性真菌病诊断方法的应用人群、送检要求、结果解读及临床意义进行了介绍并提出建议,强调对于侵袭性真菌病高危患者,应综合应用多种诊断方法、正确取材送检、提高结果解读能力、了解不同方法的临床意义,以便合理应用,更好地服务于患者。     

HIV感染者侵袭性真菌病的诊治进展

侵袭性真菌病是HIV/AIDS患者机会性感染和死亡的主要原因。近2年来国内外多部关于HIV/AIDS感染者侵袭性真菌病诊断与治疗的指南与共识相继发布, 真菌病原学诊断、抗原抗体检测、分子生物学检测方法均取得了新的进展。治疗方面, 国内外对于念珠菌病、隐球菌病、肺孢子菌肺炎等HIV/AIDS患者不同类型侵袭性真菌病的治疗方案求同存异, 新型抗真菌药物的研发及抗逆转录病毒疗法的优化也进一步推动了侵袭性真菌病诊疗的进步。本研究针对HIV/AIDS患者侵袭性真菌病相关的诊断及治疗进展予以综述, 以期为临床诊疗工作提供参考。     

支气管镜在侵袭性肺真菌病诊断中的应用价值

目的了解支气管镜技术在侵袭性肺真菌病(IPFD)诊断中的应用价值。方法回顾性分析2011年1月至2015年12月在北京朝阳医院住院诊断为侵袭性肺真菌病患者210例。对210例侵袭性肺真菌病病例的病史资料进行回顾性统计分析。结果侵袭性肺真菌病患者210例,其中确诊患者30例(14.3%),临床诊断患者79例(37.6%),拟诊患者101例(48.1%);男139例,女71例,平均年龄64岁。210例中行支气管镜检查患者109例,未行支气管镜患者101例,在支气管镜组确诊患者占20.2%(22/109),非支气管镜组确诊患者占14.3%(8/101)。两组的确诊率比较,支气管镜组确诊率高于非支气管镜组(P0.05)。210例患者中痰真菌涂片及真菌培养(以下简称痰检)阳性率29.8%(59/198);支气管肺泡灌洗液(BALF)真菌涂片及真菌培养阳性率7.9%(8/101);G试验阳性率13.3%(22/166);支气管黏膜活检(EBB)阳性率22.1%(15/68);经皮肺穿刺活检术(PCNB)阳性率47.4%(9/19)。同时行痰检+G试验阳性率41.1%(65/158);同时行痰检+G试验+BALF+EBB检查阳性率57.1%(28/49)。痰检+G+BALF+EBB组的阳性率高于痰检+G组患者,两者差异有统计学意义(P0.05);痰检+G+BALF+EBB组的阳性率与PCNB组差异无统计学意义(P0.05)。结论支气管镜检查有助于IPFD患者确诊率的提高,联合应用包括支气管镜在内的多种检查手段有助于IPFD患者获得微生物学及组织病理学的诊断依据。     

真菌的药敏试验进展和前景

随着介入操作和免疫治疗发展、临床医师意识增强和实验室检测能力提高,侵袭性真菌病(IFD)的发病率和诊断水平均有上升。新的抗真菌药如泊沙康唑、棘白霉素类上市,给治疗带来更多选择。然而,抗真菌药物的耐药现象屡有报道。因此,临床对真菌药敏试验的需求日益增加。本期新版《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(第六次修订版)》(下称原则)增加了真菌药敏试验部分,以期为IFD的治疗提供参考依据。     

真菌体外鉴定和药敏试验的临床意义

由于免疫缺陷人群的增多，侵袭性真菌感染发病率逐渐增加，但真菌感染诊断的现状却远远不能满足临床需要。一方面是真菌早期病原学诊断方法还远不如细菌学诊断那么完善，早期诊断非常困难；另一方面现有的真菌学知识还不被广大临床医生熟知，因此很多患者病情延误导致治疗失败或失去了治疗的机会。为了更快提高真菌病的诊断和治疗水平，临床微生物实验室应密切配合临床，一方面做好真菌的常规检验，并积极开创真菌检验新方法，另一方面也有义务向临床医生介绍如何正确使用真菌学的诊断方法，如何正确解释真菌的实验室报告。作为临床医生，要在熟悉侵袭性真菌感染的临床表现、影像学特点的基础上，争取病原学诊断。用真菌学知识指导临床的诊断和治疗。     

肺部真菌病实验室诊断的临床应用

近年来,肺部真菌感染的发生率不断增加,但由于其临床表现并无特异性,临床上极易误诊和漏诊而丧失治疗时机.实验室检查是诊断肺部真菌病的关键,但确诊的依据只有病理组织学检查发现真菌侵袭的证据,或正常情况下无菌体腔标本真菌培养阳性.近来,一些新的诊断方法问世,如血清真菌抗原β-D-葡聚糖(G试验)和半乳甘露聚糖(GM)检测,极大地推动了侵袭性真菌感染的早期临床诊断,而PCR是非常有前景的真菌感染诊断方法.然而,最基本、最常用的依然是痰、血、经支气管镜采集标本的检测和分析.但需要规范采集标本的方法,对其结果需要结合临床谨慎地进行评估和判断.     

规范侵袭性肺真菌病的诊断与治疗

随着免疫功能受损宿主的增加、抗生素和免疫抑制剂及其在侵袭性诊疗技术的广泛使用,肺真菌病日渐增多,引起广泛关注.然而在临床上对肺真菌病的认识仍存在很多问题,一方面,对肺真菌病的警惕性不高、认识不足,常出现漏诊误诊,错失治疗机会;另一方面,由于肺真菌病的病原体绝大多数都是条件致病菌,痰标本分离出的真菌并不足于诊断,所以临床上又常出现过诊过治现象.在各类肺真菌病中,侵袭性肺真菌病病情最严重,病死率最高.因此,有必要规范侵袭性肺真菌病的诊断与治疗.     

米卡芬净治疗恶性血液病合并侵袭性真菌病的疗效分析

目的观查注射用米卡芬净钠治疗恶性血液病合并侵袭性真菌病的疗效。方法2007年2月至10月对福建医科大学附属协和医院福建省血液病研究所12例恶性血液病合并侵袭性真菌病患者,应用注射用米卡芬净钠,剂量100mg/d,分析其疗效、起效时间及不良反应发生率。结果临床总有效率为66.7%,临床诊断病例与拟诊病例有效率分别为57.14%(4/7)和80%(4/5);临床诊断病例与拟诊病例有效率相比,差异无显著性意义;注射用米卡芬净钠起效时间1~5d。结论注射用米卡芬净钠疗效显著,不良反应少,对恶性血液病合并侵袭性真菌病患者疗效较好。     

侵袭性肺真菌病14例确诊病例临床特点分析

临床上肺部真菌病早期诊断困难,其病死率高达30%～80%[1].真菌感染的诊断标准主要依据痰标本真菌培养阳性,但因上呼吸道念珠菌和曲霉污染和定植十分常见,因此早期进行肺活检可提高侵袭性肺真菌病的确诊率.     

侵袭性肺真菌病诊治的再认识

近年来,侵袭性肺真菌病(invasive pulmonary fungal diseases,IPFD)已经逐渐为广大医务工作者所认识,尤其是国内外各种有关侵袭性真菌感染的诊治指南不断问世,如欧洲癌症研究和治疗组织/侵袭性真菌感染协作组(EORTC/IFICG)和美国真菌病研究组(MSG)制定的共识[1],美国感染病学会(IDSA)制定的曲霉病和念珠菌病临床实践指南[2-3],我国血液、呼吸和ICU等专业委员会制定的有关侵袭性真菌感染(IFI)的诊治原则以及中华医学会呼吸病学分会制定的"肺真菌病诊断和治疗专家共识"[4]等,极大地提高了临床医生对侵袭性真菌病的认识和诊治水平,但同时我们在临床工作中也发现,即使按照共识或指南的要求去做,依然还有许多问题不能解决,如早期诊断的可靠性、生物标志物的临床诊断价值、经验性治疗的时机和疗程等.随着近年来循证医学证据的不断增多,这些问题逐渐有了新的解释和认识,现结合自己的临床诊治体会谈点浅知拙见,供大家参考指正.     

A consensus document for the clinical management of invasive fungal diseases in pediatric patients with hematologic cancer and/or undergoing hematopoietic stem cell transplantation in Brazilian medical centers

In the present paper we summarize the suggestions of a multidisciplinary group including experts in pediatric oncology and infectious diseases who reviewed the medical literature to elaborate a consensus document (CD) for the diagnosis and clinical management of invasive fungal diseases (IFDs) in children with hematologic cancer and those who underwent hematopoietic stem-cell transplantation. All major multicenter studies designed to characterize the epidemiology of IFDs in children with cancer, as well as all randomized clinical trials addressing empirical and targeted antifungal therapy were reviewed. In the absence of randomized clinical trials, the best evidence available to support the recommendations were selected. Algorithms for early diagnosis and best clinical management of IFDs are also presented. This document summarizes practical recommendations that will certainly help pediatricians to best treat their patients suffering of invasive fungal diseases.     

Impact of diagnostic markers on early antifungal therapy

PURPOSE OF REVIEW: The early treatment of invasive fungal infection is critical but is hampered by the non-specific nature of clinical and radiological signs and the insensitivity of current laboratory diagnostic methods. If mortality due to invasive fungal infection is to be reduced, new, preemptive therapeutic strategies, targeting those patients at highest risk, are required and these will depend on the development of rapid, sensitive diagnostic methods. Such methods have become available in the form of high-resolution computed tomography scanning and serological and molecular techniques and in this review the authors describe recent studies assessing the utility of these methods and consider their role in management strategies. RECENT FINDINGS: Sensitive assays for the detection of fungal DNA and antigens such as galactomannan and glucan have been prospectively evaluated in the clinical setting and enable identification of patients at an earlier stage of infection. However, the sensitivity and specificity of the assays vary considerably in different studies, depending on several factors including patient selection and clinical application of the test, and issues regarding the release and circulation of galactomannan and fungal DNA remain to be clarified. SUMMARY: Rapid serological and molecular diagnostic methods facilitate the early diagnosis of invasive fungal infection and would appear to be most useful when used prospectively to screen high-risk patients. However, in order to determine the optimal approach to treatment it is essential that these tests are incorporated into management strategies and their impact on incidence of invasive fungal infection and clinical outcome evaluated in further clinical trials.     

Enfermedad fúngica invasora: ¿Diagnóstico micológico convencional o molecular?

Diagnosis of invasive mycoses is a difficult challenge due to the limitations and low sensitivity of traditional microbiology methods which lead to diagnostic and therapeutic delays. The aim of this review is to summarise the state of the art of the molecular diagnosis of invasive fungal disease and to clarify its current role in the clinical practice. Conventional microbiological methods could be complemented with molecular methods in the rapid and definitive identification of fungal isolates. Biomarkers (β-glucan, galactomannan) are very useful in immunocompromised patients and have been included as probable invasive mycoses by the EORTC/MSG. Nucleic acid detection is currently used as a complementary tool for diagnosis. However, PCR can be very useful in mould invasive mycoses. Finally, the combined detection using biomarkers can improve the diagnosis. However, their applicability in the microbiology laboratory is not so easy and further studies are required for the appropriate evaluation of its clinical usefulness.     

侵袭性肺部真菌感染的实验室检测方法

近年来,侵袭性肺部真菌感染的发病率逐年上升,并逐渐成为导致患者死亡的重要原因之一。由于侵袭性肺部真菌感染临床表现缺乏特异性,早期诊断困难,且病情易被基础病掩盖,造成误诊﹑漏诊而延误治疗。精准的实验室诊断是实现精准临床诊疗的前提和基础。该文对目前侵袭性肺部真菌感染常用实验室检测方法,包括标本直接镜检、组织病理学检查、真菌培养、抗原检测、核酸检测等的诊断性能及注意事项进行介绍与评价,从而提高临床医师及实验室人员对侵袭性肺部真菌感染实验室检测方法的认识。     

Comparison of invasive fungal diseases between patients with acute myeloid leukemia receiving posaconazole prophylaxis and those not receiving prophylaxis: A single-center,observational, case-control study in South Korea

Posaconazole prophylaxis is effective in decreasing the incidence of invasive fungal diseases (IFDs) in patients with acute myeloid leukemia (AML). However, the use of antifungal prophylaxis varies in real-life practice, and only a small number of studies have compared the incidence of IFDs between those receiving posaconazole prophylaxis and those without prophylaxis. We compared the clinical characteristics and outcomes of IFDs between patients with AML who received posaconazole prophylaxis and those without antifungal prophylaxis.We reviewed the medical records of adult AML patients who underwent induction chemotherapy between June 2016 and October 2019 at Asan Medical Center (Seoul, South Korea), where posaconazole prophylaxis is not administered in patients with gastrointestinal symptoms that may hinder sufficient absorption of oral prophylactic agents, and in patients with abnormal liver functions considering the possible exacerbation of adverse events. Patients who received posaconazole prophylaxis for ≥7 days were included in the prophylaxis group. Clinical characteristics and outcomes including the incidence of IFDs were compared between the 2 groups.Of the 247 patients with AML who underwent induction chemotherapy, 162 (66%) received posaconazole prophylaxis and 85 (34%) did not receive any prophylaxis. The incidence of proven/probable IFD was significantly higher in the no prophylaxis group than in the prophylaxis group (9.4% [8/85] vs 2.5% [4/162], P = .03). Of the 8 cases of IFDs in the no prophylaxis group, 7 were mold infections and 1 was invasive candidiasis. Of the 4 cases of IFDs in the prophylaxis group, 3 were mold infections and 1 was invasive candidiasis. Patients with posaconazole prophylaxis less frequently received therapeutic antifungal therapy (2.5% vs 9.4%, P = .03) and had a longer median, duration from chemotherapy to antifungal therapy compared with the no prophylaxis group (18 vs 11 days, P < .01). The rate of IFD-related mortality was similar between the 2 groups (0.6% vs 0%, P > .99).Patients with AML who received posaconazole prophylaxis had a lower incidence of breakthrough IFDs compared with those who did not receive any prophylaxis. Invasive mold infection was the most common IFD regardless of antifungal prophylaxis.     

Advances in the molecular and serological diagnosis of invasive fungal infection in haemato-oncology patients

Current laboratory diagnostic methods for invasive fungal infection (IFI) in haemato-oncology patients are insensitive, resulting in late diagnosis and contributing to high mortality. In recent years, progress has been made in the development and evaluation of sensitive sero-diagnostic assays, including detection of genomic DNA sequences and fungal antigens, which aid in a rapid, early diagnosis of IFI. The sensitivity and specificity of the assays vary considerably between studies, highlighting the need to correlate serological results with conventional laboratory tests and clinical or radiological findings. As part of management protocols, these assays may help to confirm the diagnosis of suspected IFI; however, the impact on mortality from IFI may be greatest when they are used to screen high-risk patients. Persistently positive screening results could direct early aggressive antifungal therapy, guided further by radiological and microbiological findings combined with regular clinical review, while the excellent negative predictive value may allow treatment to be withheld in patients with antibiotic resistant neutropenic fever but no other signs of IFI. However, this pre-emptive approach requires evaluation in prospective randomized trials.     

肝衰竭合并侵袭性真菌病的诊断策略

肝衰竭患者发生侵袭性真菌病(IFD)的危险性增加,致死率增加,早期诊断、及时治疗是改善预后的关键。笔者梳理了肝衰竭合并IFD的流行情况及危险因素、肝衰竭患者的早期识别、肝衰竭合并IFD的诊断、中医对肝衰竭合并真菌感染的认识等内容。肝衰竭患者凝血机制差,获取病理确诊较为困难,详细分析患者的危险因素、早期识别临床特征、及时进行血液/体液微生物学检测是早期诊断的重要策略。     

Early diagnosis of invasive fungal infections in hematooncological patients by serological methods

Invasive fungal infections have become one of the major causes of morbidity and mortality in hematooncological patients over the past 2 decades. Even there is an increasing incidence of invasive fungal infections caused by rare filamentous fungi, the majority of infections are caused by Candida sp. and especially Aspergillus sp. Early diagnosis and prompt initiation of antifungal treatment are leading factors influencing prognosis of patients with invasive fungal infection. Important advances in the field of early diagnosis of invasive fungal infections have been realized over the last years. Beside of new radiological methods the major progress has been done in serological methods. In this paperwe review the most important of these serological methods and their position in routine clinical practice.     

Selected risk factors of fungal infections in human populations

For the past twenty years, invasive fungal infections have dramatically increased resulting in the high morbidity and mortality. The main reason of these infections is the development of invasive diagnostic and therapeutic technologies. Immunosuppression, neutropenia, broad-spectrum antibacterial agents, corticosteroids, chemotherapy, central intravascular catheter, total parenteral nutrition are the major risk factors of the invasive fungal infections. Most of these factors are very common for hospitalized patients. For example, patients with fungemia are more likely to die during hospitalization then those with bloodstream infection caused by nonfungal pathogens. Unfortunately, diagnosis and diagnostic methods of fungal infection are still difficult.