Department of Medical Microbiology, Infection and Immunity, South West London Pathology, St George's University Hospitals NHS Foundation Trust, London, UK.
School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK.
J Med Virol. 2023 Sep;95(9):e29081. doi: 10.1002/jmv.29081.
Sample collection, transport and storage conditions vary in the human cytomegalovirus (CMV) shedding literature. Currently, limited data exist on the impact of biological fluids and pre-analytical sample handling on the detection of CMV DNA. To evaluate CMV DNA recovery from urine, vaginal fluid and saliva stored in different conditions, adult urine, vaginal and saliva fluids and swabs, stored with or without selected nucleic acid preservation media at various durations and temperatures, was compared by polymerase chain reaction (PCR) quantitation of spiked samples and self-collected urine (n = 45) and vaginal swabs (n = 58) from CMV seropositive pregnant women. There was a time-dependent reduction in CMV DNA recovery from urine, urine diluted in phosphate-buffered saline, and saliva stored at 2-8°C, but not from urine preserved in cobas® PCR transport media (CPM) (urine/CPM). For vaginal fluid, a reduction in recovery was evident after 7 days storage at 2-8°C. CMV DNA recovery over 91 days was similar between -80°C and -20°C storage for urine and vaginal swabs preserved in CPM, and saliva swabs preserved in eNAT® PCR transport media. A statistically significant change in CMV DNA recovery after 25 months storage (median) at -80°C was not observed for self-collected urine/CPM and vaginal swab/CPM from pregnant women. Taken together, recovery of CMV DNA is dependent on fluid type and storage conditions. To improve the validity and reliability of detection at different storage durations and temperatures, the use of nucleic acid preserving transport media at the point of collection for urine, vaginal fluid and saliva may be essential.
样本采集、运输和储存条件在人类巨细胞病毒（CMV）脱落文献中有所不同。目前，关于生物体液和样本前处理对 CMV DNA 检测的影响的数据有限。为了评估不同条件下储存的尿液、阴道分泌物和唾液中 CMV DNA 的恢复情况，我们通过聚合酶链反应（PCR）定量分析了加标样本和 CMV 血清阳性孕妇自行采集的尿液（n=45）和阴道拭子（n=58），比较了在不同时间和温度下，将成人尿液、阴道和唾液样本与或不与选定的核酸保存介质一起储存的情况。结果显示，尿液、磷酸盐缓冲盐水稀释的尿液和 2-8°C 储存的唾液中 CMV DNA 的恢复量随时间呈下降趋势，但尿液保存在 cobas® PCR 运输介质（CPM）（尿液/CPM）中则不然。对于阴道分泌物，2-8°C 储存 7 天后，恢复量明显减少。尿液和阴道拭子保存在 CPM 中，-80°C 和 -20°C 储存 91 天以上，唾液拭子保存在 eNAT® PCR 运输介质中，CMV DNA 的恢复情况相似。在 -80°C 储存 25 个月（中位数）后，未观察到孕妇自行采集的尿液/CPM 和阴道拭子/CPM 中 CMV DNA 恢复情况有统计学意义的变化。综上所述，CMV DNA 的恢复情况取决于体液类型和储存条件。为了提高不同储存时间和温度下检测的有效性和可靠性，在采集时使用核酸保存运输介质对尿液、阴道液和唾液可能是必要的。
