Department of Clinical Chemistry, University of Medicine in Wroclaw, Wroclaw, Poland.
Med Sci Monit. 2012 Jun;18(6):BR221-8. doi: 10.12659/msm.882898.
Detection and quantification of adenoviruses (AdVs) causing life-threatening complications are important abilities in recognition of infection and management of immunocompromised patients. Due to the rapid increase in the number of known AdV types, most commercial tests for detection and identification of AdVs are outdated.
MATERIAL/METHODS: We designed an improved, easier and faster real-time quantitative polymerase chain reaction (RQ-PCR) method for detection and quantification of 54 types of human AdVs. A wide validation effort was undertaken to ensure confidence in highly sensitive and specific detection of AdVs in compromised patients. The validation process included evaluation of the method's suitability and reliability for use in routine diagnostics.
Due to high sensitivity (9.2×10² copies/ml) and broad dynamic range (7 log) we are able to detect specific viral DNA in large amounts of cell-free body fluids. The new assay is characterized by high precision and low variation within and between individual virus tests (CV=0.036%, CV=1.29%), low bias error (4%) and no cross-reactivity with other pathogens.
The implementation of this new assay in clinical and laboratory practice provides a rapid, reliable and less laborious method for detection and monitoring of AdV replication in immunocompromised patients. Moreover, it offers the ability to distinguish between active and latent infection and assess treatment efficiency.
检测和定量引发危及生命并发症的腺病毒（AdV），对于识别感染和管理免疫功能低下患者至关重要。由于已知 AdV 类型数量的快速增加，大多数用于检测和鉴定 AdV 的商业检测方法已经过时。
材料/方法：我们设计了一种改进的、更简单和更快的实时定量聚合酶链反应（RQ-PCR）方法，用于检测和定量 54 种人类 AdV。进行了广泛的验证工作，以确保在免疫功能低下患者中高度敏感和特异性检测 AdV 的信心。验证过程包括评估该方法在常规诊断中的适用性和可靠性。
由于高灵敏度（9.2×10² 拷贝/ml）和宽动态范围（7 对数），我们能够在大量无细胞体液中检测到特定的病毒 DNA。新的检测方法具有高精度和个体病毒检测内和之间的低变异（CV=0.036%，CV=1.29%）、低偏差误差（4%）和与其他病原体无交叉反应性的特点。
在临床和实验室实践中实施这种新的检测方法为免疫功能低下患者中 AdV 复制的检测和监测提供了一种快速、可靠和不那么繁琐的方法。此外，它还能够区分活动性和潜伏性感染，并评估治疗效果。
