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Abstract：
Invasive aspergillosis (IA) by a triazole-resistant Aspergillus fumigatus is associated with high mortality. Real-time resistance detection will result in earlier initiation of appropriate therapy.
In a prospective study, we evaluated the clinical value of the AsperGenius polymerase chain reaction (PCR) assay in hematology patients from 12 centers. This PCR assay detects the most frequent cyp51A mutations in A. fumigatus conferring azole resistance. Patients were included when a computed tomography scan showed a pulmonary infiltrate and bronchoalveolar fluid (BALf) sampling was performed. The primary end point was antifungal treatment failure in patients with azole-resistant IA.
Of 323 patients enrolled, complete mycological and radiological information was available for 276 (94%), and probable IA was diagnosed in 99/276 (36%). Sufficient BALf for PCR testing was available for 293/323 (91%). Aspergillus DNA was detected in 116/293 (40%) and A. fumigatus DNA in 89/293 (30%). The resistance PCR was conclusive in 58/89 (65%) and resistance detected in 8/58 (14%). Two had a mixed azole-susceptible/azole-resistant infection. In the 6 remaining patients, treatment failure was observed in 1. Galactomannan positivity was associated with mortality (P = .004) while an isolated positive Aspergillus PCR was not (P = .83).
Real-time PCR-based resistance testing may help to limit the clinical impact of triazole resistance. In contrast, the clinical impact of an isolated positive Aspergillus PCR on BALf seems limited. The interpretation of the EORTC/MSGERC PCR criterion for BALf may need further specification (eg, minimum cycle threshold value and/or PCR positive on >1 BALf sample).
In a prospective study, we evaluated the clinical value of the AsperGenius polymerase chain reaction (PCR) assay in hematology patients from 12 centers. This PCR assay detects the most frequent cyp51A mutations in A. fumigatus conferring azole resistance. Patients were included when a computed tomography scan showed a pulmonary infiltrate and bronchoalveolar fluid (BALf) sampling was performed. The primary end point was antifungal treatment failure in patients with azole-resistant IA.
Of 323 patients enrolled, complete mycological and radiological information was available for 276 (94%), and probable IA was diagnosed in 99/276 (36%). Sufficient BALf for PCR testing was available for 293/323 (91%). Aspergillus DNA was detected in 116/293 (40%) and A. fumigatus DNA in 89/293 (30%). The resistance PCR was conclusive in 58/89 (65%) and resistance detected in 8/58 (14%). Two had a mixed azole-susceptible/azole-resistant infection. In the 6 remaining patients, treatment failure was observed in 1. Galactomannan positivity was associated with mortality (P = .004) while an isolated positive Aspergillus PCR was not (P = .83).
Real-time PCR-based resistance testing may help to limit the clinical impact of triazole resistance. In contrast, the clinical impact of an isolated positive Aspergillus PCR on BALf seems limited. The interpretation of the EORTC/MSGERC PCR criterion for BALf may need further specification (eg, minimum cycle threshold value and/or PCR positive on >1 BALf sample).
摘要：
背景：由三唑抗性烟曲霉引起的侵袭性曲霉病（IA）与高死亡率相关。实时抗性检测将导致更早开始适当的治疗。
方法：在荷兰和比利时的一项前瞻性研究中，我们评估了AsperGenius®多重PCR在12个中心血液病患者中的临床价值.该PCR检测赋予唑抗性的烟曲霉中最常见的cyp51A突变。当CT扫描显示肺部浸润并进行支气管肺泡灌洗（BALf）采样时，将患者包括在内。主要终点是唑类药物耐药IA患者的抗真菌治疗失败。排除混合唑敏感/耐药感染的患者。
结果：在323名患者中，276/323(94%)患者可获得完整的真菌学和放射学信息,99/276(36%)患者可获得可能的IA.用于PCR测试的足够的BALf在293/323(91%)中可用。在116/293(40%)中检测到曲霉DNA,在89/293(30%)中检测到烟曲霉DNA。抗性PCR在58/89(65%)中是决定性的,并且在8/58(14%)中检测到抗性。两个患有混合的唑敏感/耐药感染。在剩下的6名患者中，在一个患者中观察到治疗失败.半乳甘露聚糖阳性与较高的死亡率相关(p=0.004)。相比之下,分离的曲霉属PCR阳性患者的死亡率与PCR阴性患者的死亡率相当(p=0.83).
结论：基于实时PCR的耐药性检测可能有助于限制三唑耐药性的临床影响。相比之下,分离的曲霉PCR阳性对BALf的临床影响似乎有限.对BALf的EORTC/MSGERCPCR标准的解释可能需要进一步说明（例如，最小Ct值和/或>1个BALf样品上的PCR阳性）。
方法：在荷兰和比利时的一项前瞻性研究中，我们评估了AsperGenius®多重PCR在12个中心血液病患者中的临床价值.该PCR检测赋予唑抗性的烟曲霉中最常见的cyp51A突变。当CT扫描显示肺部浸润并进行支气管肺泡灌洗（BALf）采样时，将患者包括在内。主要终点是唑类药物耐药IA患者的抗真菌治疗失败。排除混合唑敏感/耐药感染的患者。
结果：在323名患者中，276/323(94%)患者可获得完整的真菌学和放射学信息,99/276(36%)患者可获得可能的IA.用于PCR测试的足够的BALf在293/323(91%)中可用。在116/293(40%)中检测到曲霉DNA,在89/293(30%)中检测到烟曲霉DNA。抗性PCR在58/89(65%)中是决定性的,并且在8/58(14%)中检测到抗性。两个患有混合的唑敏感/耐药感染。在剩下的6名患者中，在一个患者中观察到治疗失败.半乳甘露聚糖阳性与较高的死亡率相关(p=0.004)。相比之下,分离的曲霉属PCR阳性患者的死亡率与PCR阴性患者的死亡率相当(p=0.83).
结论：基于实时PCR的耐药性检测可能有助于限制三唑耐药性的临床影响。相比之下,分离的曲霉PCR阳性对BALf的临床影响似乎有限.对BALf的EORTC/MSGERCPCR标准的解释可能需要进一步说明（例如，最小Ct值和/或>1个BALf样品上的PCR阳性）。
