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高白细胞计数和氢化可的松的使用会干扰基于 PCR 的血流感染诊断。

A high leukocyte count and administration of hydrocortisone hamper PCR-based diagnostics for bloodstream infections.

机构信息

Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, Schöpfstrasse 41, 6020, Innsbruck, Austria.

Ares Genetics GmbH, Karl-Farkas-Gasse 18, 1030, Vienna, Austria.

出版信息

Eur J Clin Microbiol Infect Dis. 2021 Jul;40(7):1441-1449. doi: 10.1007/s10096-020-04126-w. Epub 2021 Feb 5.

Abstract

Bloodstream infections (BSIs) require an accurate and fast identification of causative pathogens. Molecular diagnostics, in particular polymerase chain reaction (PCR)-based approaches for BSI diagnostics directly from whole blood, suffer from limitations such as inhibition leading to invalid results. In this retrospective study, we analyzed 23 parameters for their potential interference with LightCycler SeptiFast PCR tests (n = 2167) routinely performed at our institution. The overall inhibition rate was 9.1%. Test date, type of ward, procalcitonin levels, high leukocyte counts, and absolute neutrophil count were significantly associated with inhibition. For a subset (n = 448), cut-off values for leukocyte counts of < 5700 cells/μL and ≥ 26,900 cells/μL were significantly associated with a low (5%) and high (67%) inhibition risk. For patients with a moderate to high leukocyte count (5700-26,900 cells/μL), the additional administration of hydrocortisone significantly increased the inhibition risk. Furthermore, freezing of blood samples prior to DNA extraction and SF testing appeared to neutralize inhibitory factors. It remains to be investigated whether other molecular diagnostic tests are susceptible to similar inhibiting parameters.

摘要

血流感染(BSI)需要准确和快速地鉴定病原体。分子诊断学,特别是聚合酶链反应(PCR)为基础的方法,直接从全血中进行 BSI 诊断,受到抑制等限制,导致结果无效。在这项回顾性研究中,我们分析了 23 个参数,以评估它们对本机构常规进行的 LightCycler SeptiFast PCR 检测(n=2167)的潜在干扰。总的抑制率为 9.1%。检测日期、病房类型、降钙素原水平、白细胞计数高和绝对中性粒细胞计数与抑制显著相关。对于一个亚组(n=448),白细胞计数<5700 个/μL 和≥26900 个/μL 的截断值与低(5%)和高(67%)抑制风险显著相关。对于白细胞计数中度至高度升高(5700-26900 个/μL)的患者,额外给予氢化可的松会显著增加抑制风险。此外,在进行 DNA 提取和 SF 检测之前冷冻血样似乎可以中和抑制因子。其他分子诊断测试是否容易受到类似的抑制参数的影响仍有待研究。

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