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使用改良的商品化方法进行全自动 D-乳酸检测。

Fully automatic d-lactate assay using a modified commercially available method.

机构信息

Department of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark.

Department of Surgery, Aalborg University Hospital, Aalborg, Denmark.

出版信息

Scand J Clin Lab Invest. 2021 Jul;81(4):312-317. doi: 10.1080/00365513.2021.1907859. Epub 2021 Apr 20.

Abstract

Intestinal infarction is the fast-evolving endpoint of impaired blood perfusion to an intestinal segment which may have fatal outcome. Early diagnosis and treatment within 6 h reduce mortality. Currently, d-lactate is a promising biomarker, however, not available in the acute clinical setting. The aim of this study is implementation of d-lactate analysis in a routine clinical setting. We used a spectrophotometric method, based on enzymatic oxidation of d-lactate by d-lactate dehydrogenase (D-LDH) coupled to the reduction of nicotinamide-adenine dinucleotide (NAD). The amount of NADH formed in this reaction is equivalent to d-lactate. The primary concern in this method is interfering NADH formed by oxidation of l-lactate by l-lactate dehydrogenase (L-LDH). A commercially available kit for d-lactate measurement was implemented on our existing automated routine laboratory equipment including pH-inactivation of L-LDH. Our setup fulfilled clinical quality goals. We were able to measure d-lactate with an acceptable performance of the analysis and a short turn-around time. The method can be used to distinguish between the expected cut-off for intestinal ischemia around 0.3 mM and the upper reference limit of 0.05 mM. With a turnaround time of just 9 min, the analysis has potential as a readily available detection of circulating d-lactate for early diagnosis of intestinal ischemia.

摘要

肠梗死是肠道某一血供受损节段的快速进展性终末事件,可能导致致命后果。6 小时内的早期诊断和治疗可降低死亡率。目前,D-乳酸是一种很有前途的生物标志物,但在急性临床环境中不可用。本研究的目的是在常规临床环境中实施 D-乳酸分析。我们使用分光光度法,基于 D-乳酸脱氢酶(D-LDH)对 D-乳酸的酶促氧化,与烟酰胺腺嘌呤二核苷酸(NAD)的还原偶联。在该反应中形成的 NADH 的量与 D-乳酸相当。该方法的主要关注点是由 L-乳酸脱氢酶(L-LDH)氧化 L-乳酸形成的干扰 NADH。我们在现有的自动化常规实验室设备上实施了一种商业上可用于 D-乳酸测量的试剂盒,包括 pH 灭活 L-LDH。我们的设置满足临床质量目标。我们能够以可接受的分析性能和较短的周转时间测量 D-乳酸。该方法可用于区分预期的肠缺血 0.3mM 左右的截止值和 0.05mM 的上限参考值。分析的周转时间仅为 9 分钟,具有作为早期诊断肠缺血的循环 D-乳酸的快速检测的潜力。

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