Biological Research and Services Laboratory, Natural Sciences Research Institute, University of the Philippines Diliman, Quezon City 1101, Philippines; Pathogen-Host-Environment Interactions Research Laboratory, Institute of Biology, College of Science, University of the Philippines Diliman, Quezon City 1101, Philippines.
Biological Research and Services Laboratory, Natural Sciences Research Institute, University of the Philippines Diliman, Quezon City 1101, Philippines.
J Virol Methods. 2024 Dec;330:115041. doi: 10.1016/j.jviromet.2024.115041. Epub 2024 Oct 9.
The human adenovirus (HAdV) has shown greater environmental persistence, more water treatment resistance than bacteria, and cause infection even at low concentrations. HAdV causes gastrointestinal illnesses and is abundant in various environmental samples such as groundwater, surface water, recreational water, and drinking water. The detection of these pathogens calls for a more practical and affordable approach. A recent technology based on the quenching of unincorporated amplification signal reporters (QUASR) was adapted for the detection of human adenoviruses. This technology allows for non-inhibitory, single-step DNA detection in a closed tube. The QUASR-(loop-mediated isothermal amplification (LAMP) assay was previously optimized and was tested for its applicability to detect enteric HAdV in two areas where people can unintentionally be exposed to contaminated water. A total of 203 water samples were collected and tested using both real-time PCR and QUASR-LAMP assays. Results showed a higher positivity rate of 78.82 % (160/203) for QUASR-LAMP compared to qPCR with only 58.62 % (119/203). The sensitivity and specificity rates for QUASR-LAMP were calculated at 86.55 % and 32.14 %, respectively, when compared to qPCR. The QUASR-LAMP assay's ability to detect target analytes even at low concentrations can be attributed to its increased diagnostic sensitivity but lower specificity since there were samples that were positive in PCR but negative in the QUASR-LAMP assay. However, this characteristic does not diminish its utility as a valuable tool for the detection of HAdV. In fact, this attribute enhances its advantages in situations with constrained space and instrumentation requirements, making it suitable for rapid surveillance of important viruses. Finally, the capacity of the QUASR-LAMP assay to differentiate between positive and negative samples at a defined endpoint is highly beneficial for laboratory technicians who possess limited molecular biology expertise or experience. The QUASR-LAMP platform has demonstrated its usefulness as a diagnostic tool for surveillance of enteric adenoviruses in water sources.
人类腺病毒 (HAdV) 具有较强的环境持久性和比细菌更强的耐水处理能力，即使在低浓度下也能引起感染。HAdV 会引起胃肠道疾病，并且在地下水、地表水、娱乐水和饮用水等各种环境样本中都很丰富。这些病原体的检测需要更实用和更经济的方法。最近，一种基于未结合扩增信号报告物淬灭 (QUASR) 的技术被用于检测人类腺病毒。该技术允许在封闭管中进行非抑制性、一步法 DNA 检测。QUASR-(环介导等温扩增 (LAMP) 检测法之前已经经过优化，并在两个人们可能无意中接触到污染水的地区进行了检测肠道 HAdV 的适用性测试。共采集 203 个水样，分别采用实时 PCR 和 QUASR-LAMP 检测。结果显示，与仅 58.62%（119/203）的 qPCR 相比，QUASR-LAMP 的阳性率更高，为 78.82%（160/203）。与 qPCR 相比，QUASR-LAMP 的灵敏度和特异性分别为 86.55%和 32.14%。QUASR-LAMP 检测低浓度目标分析物的能力归因于其更高的诊断灵敏度和较低的特异性，因为有些样本在 PCR 中呈阳性而在 QUASR-LAMP 检测中呈阴性。然而，这一特性并不会降低其作为 HAdV 检测的有用工具的实用性。事实上，在空间和仪器要求受限的情况下，这种特性增强了其优势，使其成为重要病毒快速监测的理想选择。最后，QUASR-LAMP 检测法在定义的终点区分阳性和阴性样本的能力对具有有限分子生物学专业知识或经验的实验室技术人员非常有益。QUASR-LAMP 平台已被证明可作为水污染源肠道腺病毒监测的诊断工具。
