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Abstract：
Talaromyces (Penicillium) marneffei (TM) is an important, but neglected, thermally dimorphic fungus. It is the pathogenic cause of talaromycosis, which is strongly associated with the immunodeficiency state present in individuals with advanced HIV disease. The purpose of this study was to develop a sandwich enzyme-linked immunosorbent assay (sandwich ELISA) for the detection of T. marneffei cytoplasmic yeast antigen (TM CYA) in human urine. Monoclonal antibody (MAb) 4D1 specifically binds to TM CYA. Galanthus nivalis agglutinin (GNA), a mannose -binding lectin, recognizes and binds to mannose residues of TM CYA. For the sandwich ELISA, the microplate was coated with GNA as the capturing molecule for absorbing immune complexes of MAb 4D1-TM CYA. The MAb 4D1-GNA sandwich ELISA did not detect a cross-reaction with other antigens from other fungi or bacteria. Seventy-four urine samples from patients with blood culture -confirmed talaromycosis and 229 urine samples from people without talaromycosis residing in the endemic area were subjected to the MAb 4D1-GNA sandwich ELISA. At an optical density (OD) cutoff value of 0.356, the sensitivity was 89.19% [95% confidence interval (CI): 79.80% -95.22%]; the specificity was 98.69% (95% CI: 96.22% -99.73%). The diagnostic performance of the MAb 4D1-GNA sandwich ELISA was highly consistent with those of blood culture and the Platelia Aspergillus galactomannan (GM) ELISA kit. Collectively, the MAb 4D1-GNA sandwich ELISA is a promising technique for the rapid diagnosis of T. marneffei infection, which would facilitate the early treatment of patients with talaromycosis and it may be used to monitor treatment responses.
摘要：
Talaromyces(Penicillium)marneffei(TM)是一种重要的，但被忽视了，热二态真菌。它是塔拉真菌病的致病原因，这与晚期HIV疾病患者的免疫缺陷状态密切相关。本研究的目的是开发一种夹心酶联免疫吸附测定法（夹心ELISA），用于检测人尿中的马尼菲氏杆菌细胞质酵母抗原（TMCYA）。单克隆抗体(MAb)4D1特异性结合TMCYA。鹅卵菜凝集素（GNA），一种甘露糖结合凝集素，识别并结合TMCYA的甘露糖残基。对于夹心ELISA，用GNA包被微孔板，作为吸收MAb4D1-TMCYA免疫复合物的捕获分子。MAb4D1-GNA夹心ELISA未检测到与来自其他真菌或细菌的其他抗原的交叉反应。对来自血液培养证实的talaryomcosis患者的74份尿液样本和来自流行地区没有talaryomcosis的人的229份尿液样本进行了MAb4D1-GNA夹心ELISA。在光密度（OD）截止值为0.356时，灵敏度为89.19％[95％置信区间（CI）：79.80％-95.22％]；特异性为98.69％（95％CI：96.22％-99.73％）。MAb4D1-GNA夹心ELISA的诊断性能与血液培养和双乳曲霉半乳甘露聚糖（GM）ELISA试剂盒的诊断性能高度一致。总的来说，MAb4D1-GNA夹心ELISA是一种快速诊断马尔尼菲感染的有前途的技术，这将有助于塔拉真菌病患者的早期治疗，并可用于监测治疗反应。
