为减少传染病污染而对唾液进行热处理不会影响通过放射免疫分析法分析褪黑素。

IF 8.3 1区 医学 Q1 ENDOCRINOLOGY & METABOLISM
Mark D. Salkeld, David J. Kennaway
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引用次数: 0

摘要

唾液中褪黑激素水平的测定是评估人类中枢昼夜节律时钟时间的关键方法之一,无论是在研究还是在临床环境中都是如此。唾液中的褪黑激素水平通常是在实验室环境中通过 RIA 或酶联免疫吸附试验测定的,而严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)的出现给唾液样本中褪黑激素的常规安全评估带来了严峻挑战。然而,生物液体中的 SARS-CoV-2 可在至少 55-60°C 的温度下暴露 30 分钟而失活,因此本研究的目的是评估使用 Novolytix 放射免疫分析仪 (RK-DSM2) 对唾液样本进行预处理加热步骤以测定褪黑激素的有效性。解冻根据暗光褪黑素起始采样方案收集的 40 份存档唾液样本,并将其等分到三个相同的组中--对照组(无预处理)、56°C 检测前热处理组(30 分钟)和 70°C 检测前热处理组(30 分钟)。检测前加热至 56°C 30 分钟的样本中褪黑激素浓度与未处理的对照组接近,两组样本之间的皮尔逊相关系数为 0.99(p<0.05)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Heat Treatment of Saliva to Reduce Infectious Disease Contamination Does Not Impact the Analysis of Melatonin by Radioimmunoassay

The determination of melatonin levels in saliva represents one of the key methods for assessing the timing of the central circadian clock in humans, both in research and clinical settings. Melatonin levels in saliva are typically determined in a laboratory setting by RIA or enzyme-linked immunosorbent assay and the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) presented a serious challenge to the routine, safe assessment of melatonin in saliva samples. However, SARS-CoV-2 present in biological fluids can be inactivated by exposure to temperatures of at least 55–60°C for 30 min and the aim of this study was to assess the validity of applying a pretreatment heating step to saliva samples being prepared for melatonin determination using the Novolytix Radioimmunoassay (RK-DSM2). 40 archived saliva samples collected under a Dim Light Melatonin Onset sampling protocol were thawed and aliquoted into three identical groups—Controls (no pretreatment), 56°C pre-assay heat-treatment (30 min), and 70°C pre-assay heat-treatment (30 min). Melatonin concentrations in samples that were heated to 56°C for 30 min before assaying showed close agreement with the untreated controls, with the Pearson's correlation coefficient between the two sets of samples of 0.99 (p < 0.0001) and the slope of the Deming regression analysis close to 1.0 (Y = 1.04X + 0.168). When saliva samples were pretreated to 70°C for 30 min before assaying, the subsequent melatonin determinations were still strongly correlated with the untreated controls (Pearsons correlation coefficient = 0.97 (p < 0.0001), however melatonin concentrations were consistently overestimated when compared to the untreated controls with Deming regression slope of Y = 1.26X + 0.241. These results indicate that a 56°C pretreatment step is suitable for inclusion in standard operating protocols for melatonin determinations using the Novolytix RIA, as a way of effectively minimizing the potential for accidental pathogen exposure while handling saliva samples.

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来源期刊
Journal of Pineal Research
Journal of Pineal Research 医学-内分泌学与代谢
CiteScore
17.70
自引率
4.90%
发文量
66
审稿时长
1 months
期刊介绍: The Journal of Pineal Research welcomes original scientific research on the pineal gland and melatonin in vertebrates, as well as the biological functions of melatonin in non-vertebrates, plants, and microorganisms. Criteria for publication include scientific importance, novelty, timeliness, and clarity of presentation. The journal considers experimental data that challenge current thinking and welcomes case reports contributing to understanding the pineal gland and melatonin research. Its aim is to serve researchers in all disciplines related to the pineal gland and melatonin.
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