Sturny-Leclère Aude, Da Silva Elodie, Godoy Cassia S M, Soares Renata B A, Leitão Terezinha do Menino Jesus Silva, Damasceno Lisandra Serra, Bay Monica B, Melo Marineide, Lana Daiane Dalla, Silva Larissa R, Israelski Dennis, Falci Diego R, Pasqualotto Alessandro C, Alanio Alexandre
Groupe de Recherche Mycologie Translationnelle, Département de Mycologie, Institut Pasteur, Université Paris Cité, Centre National de Référence Mycoses Invasives et Antifongiques, Paris, France.
Laboratoire de Parasitologie-Mycologie, AP-HP, Département de biologie médicale, Hôpital Saint-Louis, Paris, France.
Clin Microbiol Infect. 2025 Apr;31(4):636-642. doi: 10.1016/j.cmi.2024.11.037. Epub 2024 Dec 6.
Disseminated histoplasmosis (DH) is a lethal fungal disease in patients living with HIV in endemic regions of the world. Diagnosis relies mainly on microscopy, culture, and antigen detection. Our goal was to evaluate the diagnostic and prognostic role of our RT-quantitative PCR (RT-qPCR) in blood specimens allowing to quantify the whole nucleic acids (WNA) load.
We tested RT-qPCR assays on serial blood (n = 325) of 105 DH patients at baseline (day 0 or day 1, D0/1) and during antifungal therapy (day 3, D3; day 4 or day 5, D4/5; day 7, D7; and day 14, D14) collected from a phase II trial comparing three different regimens of liposomal amphotericin B for DH treatment in HIV patients from Brazil.
The RT-qPCR was positive at D0/D1 in 64.2% (63/98) of the patients, but positivity increased to 89% (47/53) in patients with proven infection. Urine antigen was positive in 94.2% (97/103) of DH patients. RT-qPCR positive at or after D7 was significantly associated with higher initial WNA load (Cq = 31 [27-34]) when compared with RT-qPCR negative at D7 (Cq = 38 [33-40]) (p 0.001). The WNA load was equivalent in all treatment arms, and an equivalent decrease in the % of RT-qPCR-positive patients was observed in the three arms. Flat of negative WNA slope (increase of WNA load) was associated with an increased relative risk of death at D7 (relative risk, 8.6; p 0.046) and W12 (relative risk, 3.3; p 0.008). This association was not observed when analysing antigen slopes (p > 0.5).
We found in this study an association between the progression of flat Histoplasma WNA load during treatment and early death at D7 and D14. This diagnostic tool should be evaluated specifically in a prospective trial to assess its usefulness in identifying patients with poorer prognosis and adapt their treatment accordingly.
播散性组织胞浆菌病（DH）是世界流行地区HIV感染者中的一种致命性真菌疾病。诊断主要依靠显微镜检查、培养和抗原检测。我们的目标是评估实时定量聚合酶链反应（RT-qPCR）在血液标本中的诊断和预后作用，该方法可定量全核酸（WNA）载量。
我们对105例DH患者在基线期（第0天或第1天，D0/1）以及抗真菌治疗期间（第3天，D3；第4或5天，D4/5；第7天，D7；第14天，D14）的系列血液样本（n = 325）进行了RT-qPCR检测，这些样本来自一项比较三种不同脂质体两性霉素B治疗方案对巴西HIV患者DH疗效的II期试验。
64.2%（63/98）的患者在D0/D1时RT-qPCR呈阳性，但确诊感染患者的阳性率增至89%（47/53）。94.2%（97/103）的DH患者尿抗原呈阳性。与D7时RT-qPCR阴性（Cq = 38 [33 - 40]）相比，D7及之后RT-qPCR呈阳性与更高的初始WNA载量（Cq = 31 [27 - 34]）显著相关（p < 0.001）。所有治疗组的WNA载量相当，且三个治疗组中RT-qPCR阳性患者百分比的下降幅度相当。WNA斜率为负（WNA载量增加）与D7时死亡相对风险增加（相对风险，8.6；p < 0.046）以及W12时死亡相对风险增加（相对风险，3.3；p < 0.008）相关。分析抗原斜率时未观察到这种相关性（p > 0.5）。
我们在本研究中发现，治疗期间组织胞浆菌WNA载量呈平缓趋势与D7和D14时早期死亡之间存在关联。应在前瞻性试验中专门评估这种诊断工具，以评估其在识别预后较差患者并据此调整治疗方面的实用性。
