Table_1_Candida-Reactive T Cells for the Diagnosis of Invasive Candida Infection—A Prospective Pilot Study.DOCX

Background: Blood or tissue culture or histology prove invasive Candida infection, but long time to result, limited feasibility and sensitivity call for new approaches. In this pilot project, we describe the diagnostic potential of quantitating Candida-reactive, CD4/CD69/CD154 positive lymphocytes in blood of patients with invasive Candida infection.
Methods: We used flow cytometry quantitating Candida-reactive, CD4/CD69/CD154 positive lymphocytes from peripheral blood of patients with invasive Candida infection, from patients at risk and healthy volunteers as controls.
Results: Elevated levels of Candida-reactive lymphocytes were measured in 13 patients with proven invasive Candida infection and in one patient with probable hepatosplenic candidiasis. Results of three candidemia patients were uninterpretable due to autofluorescence of samples. Twelve of 13 patients had Candida identified to species level by conventional methods, and T cell reactivity correctly identified Candida species in 10 of 12 patients. Nine hematological high-risk patients and 14 healthy donors had no elevated Candida-reactive T cell counts.
Conclusions: This Candida-reactive lymphocyte assay correctly identified the majority of patients with invasive Candida infection and the respective species. Our assay has the potential to support diagnosis of invasive Candida infection to species level and to facilitate tailored treatment even when biopsies are contraindicated or cultures remain negative.

背景：目前临床确诊侵袭性念珠菌感染主要依靠血液培养、组织培养或组织病理学检测，但此类检测方法存在结果回报周期长、应用可行性受限且灵敏度不足等缺陷，亟需开发新型检测策略。本预实验旨在探讨侵袭性念珠菌感染患者外周血中念珠菌反应性CD4⁺CD69⁺CD154⁺淋巴细胞定量检测的诊断价值。
方法：本研究采用流式细胞术（flow cytometry），对侵袭性念珠菌感染患者、感染高危患者以及健康志愿者（对照组）的外周血样本中的念珠菌反应性CD4⁺CD69⁺CD154⁺淋巴细胞进行定量检测。
结果：本研究中，13例经确诊的侵袭性念珠菌感染患者与1例疑似肝脾念珠菌病（hepatosplenic candidiasis）患者的外周血念珠菌反应性淋巴细胞水平均显著升高。3例念珠菌血症（candidemia）患者因样本自发荧光（autofluorescence）导致检测结果无法解读。13例确诊患者中，12例经传统检测方法明确了感染的念珠菌菌种；其中10例患者的T细胞反应性检测可正确鉴定其感染的念珠菌菌种。9例血液系统高危患者与14例健康志愿者的外周血念珠菌反应性T细胞计数均未出现升高。
结论：本研究所建立的念珠菌反应性淋巴细胞检测方法可正确识别绝大多数侵袭性念珠菌感染患者及其感染的念珠菌菌种。该检测方法有望为侵袭性念珠菌感染的菌种水平诊断提供支持，即使在活检存在禁忌或培养结果呈阴性的情况下，也可辅助临床实现个体化治疗。