唾液中腺苷脱氨酶（ADA）及其同工酶的自动化分析验证及其在 SARS-CoV-2 感染患者中变化的初步评估。

Analytical validation of an automated assay for the measurement of adenosine deaminase (ADA) and its isoenzymes in saliva and a pilot evaluation of their changes in patients with SARS-CoV-2 infection.

机构信息

Interdisciplinary Laboratory of Clinical Analysis Interlab-UMU, Regional Campus of International Excellence Mare Nostrum, University of Murcia, Espinardo, Murcia, Spain.

Preventive Medicine, Hospital Clínico Universitario Virgen de la Arrixaca, IMIB, Universidad de Murcia, Murcia, Spain.

出版信息

Clin Chem Lab Med. 2021 Apr 28;59(9):1592-1599. doi: 10.1515/cclm-2021-0324. Print 2021 Aug 26.

DOI:10.1515/cclm-2021-0324

PMID:33908223

Abstract

OBJECTIVES

The aim of the present study was to validate a commercially available automated assay for the measurement of total adenosine deaminase (tADA) and its isoenzymes (ADA1 and ADA2) in saliva in a fast and accurate way, and evaluate the possible changes of these analytes in individuals with SARS-CoV-2 infection.

METHODS

The validation, in addition to the evaluation of precision and accuracy, included the analysis of the effects of the main procedures that are currently being used for SARS-CoV-2 inactivation in saliva and a pilot study to evaluate the possible changes in salivary tADA and isoenzymes in individuals infected with SARS-CoV-2.

RESULTS

The automated assay proved to be accurate and precise, with intra- and inter-assay coefficients of variation below 8.2%, linearity under dilution linear regression with R close to 1, and recovery percentage between 80 and 120% in all cases. This assay was affected when the sample is treated with heat or SDS for virus inactivation but tolerated Triton X-100 and NP-40. Individuals with SARS-CoV-2 infection (n=71) and who recovered from infection (n=11) had higher mean values of activity of tADA and its isoenzymes than healthy individuals (n=35).

CONCLUSIONS

tADA and its isoenzymes ADA1 and ADA2 can be measured accurately and precisely in saliva samples in a rapid, economical, and reproducible way and can be analyzed after chemical inactivation with Triton X-100 and NP-40. Besides, the changes observed in tADA and isoenzymes in individuals with COVID-19 open the possibility of their potential use as non-invasive biomarkers in this disease.

摘要

目的

本研究旨在验证一种商业化的唾液总腺苷脱氨酶（tADA）及其同工酶（ADA1 和 ADA2）自动分析检测方法的准确性和精确性，评估这些分析物在 SARS-CoV-2 感染个体中的可能变化。

方法

除了评估精密度和准确性外，该验证还包括分析目前用于唾液中 SARS-CoV-2 灭活的主要步骤的影响，以及一项初步研究以评估 SARS-CoV-2 感染个体唾液中 tADA 和同工酶可能发生的变化。

结果

该自动化分析检测方法具有准确性和精确性，批内和批间变异系数均低于 8.2%，稀释线性回归的 R 值接近 1，在所有情况下回收率均在 80%至 120%之间。当样本用热或 SDS 处理以灭活病毒时，该分析检测方法会受到影响，但 Triton X-100 和 NP-40 可耐受。71 例 SARS-CoV-2 感染患者（感染恢复组）和 11 例感染恢复患者（恢复组）的 tADA 和同工酶活性的平均值均高于 35 例健康个体（对照组）。

结论

唾液中的 tADA 及其同工酶 ADA1 和 ADA2 可以通过快速、经济和可重复的方法准确、精确地测量，并且可以在 Triton X-100 和 NP-40 进行化学灭活后进行分析。此外，COVID-19 患者中观察到的 tADA 和同工酶的变化为它们在该疾病中作为非侵入性生物标志物的潜在用途提供了可能性。