新型广谱泛属 pan-genus PCR 检测方法的建立用于检测 物种。
Development of novel broad-range pan-genus PCR assays for the detection of species.
机构信息
Department of Laboratory Medicine and Pathology, Institute of Microbiology, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
Division of Immunology and Allergology, Department of Medicine, Lausanne University Hospital, Lausanne, Switzerland.
出版信息
J Med Microbiol. 2024 Sep;73(9). doi: 10.1099/jmm.0.001889.
is responsible for the classical Whipple's disease. Recently, a new species was described in a Belgian immunocompromised patient with pleuritis. There is currently no specific molecular diagnostic test detecting other species than . To develop and validate two broad-range pan- genus PCRs detecting both and new species. From shotgun sequencing data of the lung tissue biopsy of the Belgian subject, we designed two PCRs targeting the 23S rRNA and genes. Prospectively, requests for PCR were tested with -specific PCRs and the two broad-range PCRs from January 2019 to November 2022. In total, 2605 samples were tested using both the pan- 23S rRNA PCR and the -specific PCR. In addition, 833 of the 2605 samples were also tested using the pan- PCRs. Sensitivity was 78.8% and 79.7% for 23S rRNA and PCRs, as compared with the species-specific PCR. Specificity was 99.9% and 99.7% for the 23S rRNA and the PCRs, respectively. We identified a patient whose bronchoalveolar lavage tested positive with the two broad-range PCRs with >10 copies ml. Specific PCRs were negative. Known for panuveitis, this 49-year-old male presented with an eye inflammation recurrence, and a CT scan showed multiple mediastino-hilar necrotic adenopathies. Doxycyclin (1 year), hydroxychloroquin (1 year) and co-trimoxazol (1 month) treatments led to a favourable outcome. Specific PCR exhibited better sensitivity than the pan- PCRs. However, both broad-range pan- PCRs demonstrated excellent specificity and were pivotal to identifying a new probable case of infection due to another species-level lineage.
负责经典的惠普尔氏病。最近，在一名患有胸膜炎的比利时免疫功能低下的患者中描述了一个新物种。目前没有专门的分子诊断测试来检测除 以外的其他物种。开发和验证两种广泛的通用属 PCR，以检测 和新物种。从比利时患者肺部组织活检的鸟枪法测序数据中，我们设计了两种针对 23S rRNA 和 基因的 PCR。前瞻性地，从 2019 年 1 月至 2022 年 11 月，用 -特异性 PCR 对 请求的 PCR 进行了测试，以及两种 广泛的 PCR。总共使用通用 23S rRNA PCR 和 -特异性 PCR 测试了 2605 个样本。此外，833 个 2605 个样本也使用通用 PCR 进行了测试。23S rRNA 和 PCR 的敏感性分别为 78.8%和 79.7%，与物种特异性 PCR 相比。23S rRNA 和 PCR 的特异性分别为 99.9%和 99.7%。我们发现了一名患者，其支气管肺泡灌洗液用两种通用 PCR 检测呈阳性，拷贝数大于 10 拷贝/ml。特异性 PCR 为阴性。已知的全葡萄膜炎，这名 49 岁男性出现眼部炎症复发，CT 扫描显示多个纵隔-肺门坏死性淋巴结炎。多西环素（1 年）、羟氯喹（1 年）和复方磺胺甲噁唑（1 个月）治疗取得了良好的结果。特异性 PCR 显示出比通用 PCR 更高的敏感性。然而，两种广泛的通用属 PCR 都表现出了极好的特异性，对识别另一种种系的可能的 感染新病例至关重要。
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