Tamai Shinsuke, Sone Ryota, Watanabe Koichi, Shimizu Kazuhiro
Department of Sports Sciences, Japan Institute of Sports Sciences, Kita-ku 115-0056, Tokyo, Japan.
Department of Clinical Medicine, Institute of Medicine, University of Tsukuba, Tsukuba 305-8577, Ibaraki, Japan.
Viruses. 2025 Mar 14;17(3):411. doi: 10.3390/v17030411.
Salivary human herpesvirus 6 and/or 7 (HHV-6/7) have recently attracted attention as microbiological markers of physiological fatigue in laborers and athletes. However, the accuracy and efficiency of the HHV-6/7 assays can be improved for practical application. We conducted three experiments to identify suitable saliva collection and DNA extraction methods for practical salivary HHV-6/7 assays. The main experiment compared the data, including template DNA or HHV-6/7 concentrations, among three saliva collection methods (cotton, synthetic, and no swabs) and two DNA extraction methods (magnetic bead-based and silica column-based). It showed that using swabs had adverse effects: lower template DNA concentration, lower HHV-6/7 detection rates, higher coefficient of variation values, and lower concentrations. Moreover, magnetic bead-based methods resulted in higher HHV-6/7 detection rates and lower coefficient of variation values. Sub-experiment 1 examined practical saliva collection methods and demonstrated that the stimulated spitting method could collect saliva in a shorter time with lower subjective stress than the unstimulated spitting and stimulated swabbing methods. Sub-experiment 2 investigated diurnal variation in salivary HHV-6/7 levels but did not show diurnal variation. These findings suggest that (1) the combination of stimulated spitting saliva collection and magnetic bead-based DNA extraction is most suitable for practical salivary HHV-6/7 assays, and (2) saliva collection can be conducted whenever needed.
唾液人疱疹病毒6型和/或7型（HHV-6/7）最近作为劳动者和运动员生理疲劳的微生物标志物受到关注。然而，HHV-6/7检测方法的准确性和效率可针对实际应用加以提高。我们进行了三项实验，以确定适用于实际唾液HHV-6/7检测的唾液采集和DNA提取方法。主要实验比较了三种唾液采集方法（棉拭子、合成拭子和无拭子）和两种DNA提取方法（磁珠法和硅胶柱法）的数据，包括模板DNA或HHV-6/7浓度。结果表明，使用拭子有不良影响：模板DNA浓度较低、HHV-6/7检测率较低、变异系数值较高且浓度较低。此外，磁珠法导致HHV-6/7检测率较高且变异系数值较低。子实验1研究了实际唾液采集方法，结果表明，与非刺激吐唾法和刺激拭子法相比，刺激吐唾法能在更短时间内采集唾液，且主观压力较小。子实验2调查了唾液HHV-6/7水平的日变化，但未发现日变化。这些发现表明：（1）刺激吐唾唾液采集与磁珠法DNA提取相结合最适合实际唾液HHV-6/7检测；（2）可在需要时随时进行唾液采集。
