Atkinson Kelly R, Lo Kim R, Payne Steve R, Mitchell John S, Ingram John R
Functional Foods and Health Programme,The Horticultural and Food Research Institute of New Zealand Ltd., Auckland, New Zealand.
J Clin Lab Anal. 2008;22(6):395-402. doi: 10.1002/jcla.20281.
Point-of-care (POC) measurements using saliva samples have immense potential to assess systemic health and wellbeing, but sample viscosity and contaminants can affect analyses. We sought a portable clean-up method for whole saliva appropriate for use with POC measurement techniques such as biosensors.
Whole saliva from each of 13 male subjects was split into 5 fractions. Each fraction was treated with a different clean-up process: a freeze-thaw-centrifuge (FTC) step; centrifugation alone; or passage through a Mini-UniPrep polyethersulfone filter, cotton Salivette, or foam Oracol device. Following clean-up, each subject's treated saliva fractions were assayed for cortisol, testosterone, dehydroepiandrosterone (DHEA), and protein concentrations. The effects of clean-up methods on nonspecific binding (NSB) in a biosensor were also assessed.
Compared with FTC, no analytes were affected by centrifugation alone. Cotton Salivettes significantly altered all analytes, with increases in cortisol (+64%), testosterone (+126%), and DHEA (off-scale) levels, and decreased protein (-21%) and biosensor NSB (-75%). Oracol foam devices decreased DHEA levels by 28%. Mini-UniPrep filtration decreased testosterone (-45%) and DHEA (-66%) concentrations while increasing cortisol (+40%).
No method was optimal for all analytes, highlighting the need for validation of saliva treatment methods before their adoption in rapid POC analyses.
使用唾液样本进行即时检测（POC）在评估全身健康状况方面具有巨大潜力，但样本粘度和污染物会影响分析结果。我们寻求一种适用于即时检测技术（如生物传感器）的便携式全唾液净化方法。
将13名男性受试者的全唾液各分成5份。每份唾液采用不同的净化方法处理：冻融离心（FTC）步骤；仅进行离心；或通过Mini-UniPrep聚醚砜过滤器、棉质唾液采集管或泡沫Oracol装置。净化后，检测每位受试者经处理的唾液样本中的皮质醇、睾酮、脱氢表雄酮（DHEA）和蛋白质浓度。还评估了净化方法对生物传感器中非特异性结合（NSB）的影响。
与FTC相比，仅离心对所有分析物均无影响。棉质唾液采集管显著改变了所有分析物，皮质醇水平升高（+64%）、睾酮水平升高（+126%）、DHEA水平升高（超出范围），蛋白质水平降低（-21%），生物传感器NSB降低（-75%）。Oracol泡沫装置使DHEA水平降低了28%。Mini-UniPrep过滤使睾酮浓度降低（-45%）、DHEA浓度降低（-66%），同时使皮质醇浓度升高（+40%）。
没有一种方法对所有分析物都是最佳的，这突出表明在将唾液处理方法应用于快速即时检测分析之前，需要对其进行验证。
