VanDerMeid Karl R, Su Stephanie P, Krenzer Kathleen L, Ward Keith W, Zhang Jin-Zhong
Pharmaceutical R&D, Bausch + Lomb, Inc., Rochester, NY 14609, USA.
Mol Vis. 2011 Apr 27;17:1056-63.
The Schirmer's test is commonly used in the clinic for the diagnosis of dry eye disease by measuring tear volume. This report describes a procedure which can be used to recover tears from the Schirmer strip for the measurement of multiple tear cytokines as well as matrix metalloproteinases (MMPs) by Luminex technology.
Cytokine and MMP recovery was determined by using spiked Schirmer strips presoaked with known cytokines or MMPs prepared in PBS with 1% BSA. In a clinical study, tears were collected from 5 subjects using Schirmer strips. Strips were stored on ice immediately after removal from the subject and stored dry at -20 °C for 16-24 h. Cytokines were extracted from the Schirmer strip in 0.5 M NaCl with 0.5% Tween-20. Concentrations of cytokines and MMPs in collected tear samples were analyzed by Luminex using both a 10-cytokine and a 5-MMP kit.
The standard curves for the assay in both the kit assay buffer and extraction buffer were identical for 9 of the 10 cytokines and all 5 MMPs. In the clinical sample all the cytokines (interleukin 1α [IL-1α], IL-1β, IL-1ra, IL-4, IL-6, IL-8, IL-10, IL-13, monocyte chemotactic protein-1 [MCP-1], and tumor necrosis factor-α [TNF-α]) and 5 MMPs (MMP-1, MMP-2, MMP-7, MMP-9, and MMP-10) tested were detected in at least 50% of the 10 subject samples. Recoveries from extracted Schirmer strips were >60% for 8 of the 10 cytokines and all MMPs.
Numerous cytokines and MMPs were detected in the tear samples collected using the Schirmer strip, including many that have been implicated in ocular surface disease. This procedure may be used to evaluate the cytokine and MMP content in tear samples in clinical studies, especially for the evaluation of dry eye therapeutics. Because the Schirmer test is routine in the assessment of dry eye, this method offers the opportunity to evaluate both the quantity and quality of the tears.
在临床中，施密特试验常用于通过测量泪液量来诊断干眼病。本报告描述了一种可用于从施密特滤纸条回收泪液的方法，以便通过Luminex技术测量多种泪液细胞因子以及基质金属蛋白酶（MMP）。
通过使用预先浸泡有已知细胞因子或在含1%牛血清白蛋白的磷酸盐缓冲盐水中制备的MMP的加标施密特滤纸条来测定细胞因子和MMP的回收率。在一项临床研究中，使用施密特滤纸条从5名受试者收集泪液。滤纸条从受试者取出后立即保存在冰上，并在-20℃下干燥保存16 - 24小时。细胞因子从施密特滤纸条中在含0.5%吐温-20的0.5 M氯化钠中提取。使用10种细胞因子试剂盒和5种MMP试剂盒通过Luminex分析收集的泪液样本中细胞因子和MMP的浓度。
在试剂盒测定缓冲液和提取缓冲液中，10种细胞因子中的9种以及所有5种MMP的测定标准曲线相同。在临床样本中，所检测的所有细胞因子（白细胞介素1α[IL-1α]、IL-1β、IL-1ra、IL-4、IL-6、IL-8、IL-10、IL-13、单核细胞趋化蛋白-1[MCP-1]和肿瘤坏死因子-α[TNF-α]）和5种MMP（MMP-1、MMP-2、MMP-7、MMP-9和MMP-10）在10名受试者样本中至少50%被检测到。10种细胞因子中的8种以及所有MMP从提取的施密特滤纸条中的回收率>60%。
使用施密特滤纸条收集的泪液样本中检测到了多种细胞因子和MMP，包括许多与眼表疾病有关的因子。该方法可用于临床研究中评估泪液样本中的细胞因子和MMP含量，特别是用于评估干眼病治疗药物。由于施密特试验在干眼病评估中是常规操作方法，这种方法提供了评估泪液量和质的机会。
