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采用流式细胞术联合直接抗生素药敏试验快速诊断尿路感染并减少工作量。

Rapid diagnosis and reduced workload for urinary tract infection using flowcytometry combined with direct antibiotic susceptibility testing.

机构信息

Department of Microbiology, Fürst Medical Laboratory, Oslo, Norway.

出版信息

PLoS One. 2021 Jul 6;16(7):e0254064. doi: 10.1371/journal.pone.0254064. eCollection 2021.

Abstract

BACKGROUND

We evaluated if flowcytometry, using Sysmex UF-5000, could improve diagnosis of urinary tract infections by rapid identification of culture negative and contaminated samples prior to culture plating, thus reducing culture plating workload and response time. We also evaluated if it is possible to reduce the response time for antibiotic susceptibility profiles using the bacteria information flag on Sysmex UF-5000 to differentiate between Gram positive and negative bacteria, followed by direct Antibiotic Susceptibility Testing (dAST) on the positive urine samples.

METHODS

One thousand urine samples were analyzed for bacteria, white blood cells and squamous cells by flowcytometry before culture plating. Results from flowcytometric analysis at different cut-off values were compared to results of culture plating. We evaluated dAST on 100 urine samples that were analyzed as positive by flowcytometry, containing either Gram positive or Gram negative bacteria.

RESULTS

Using a cut-off value with bacterial count ≥100.000/mL and WBCs ≥10/μL, flowcytometry predicted 42,1% of samples with non-significant growth. We found that most contaminated samples contain few squamous cells. For 52/56 positive samples containing Gram negative bacteria dAST was identical to routine testing. Overall, there was concordance in 555/560 tested antibiotic combinations.

CONCLUSION

Flowcytometry offers advantages for diagnosis of urinary tract infections. Screening for negative urine samples on the day of arrival reduces culture plating and workload, and results in shorter response time for the negative samples. The bacteria information flag predicts positive samples containing Gram negative bacteria for dAST with high accuracy, thus Antibiotic Susceptibility Profile can be reported the day after arrival. For the positive samples containing Gram negative bacteria the concordance was very good between dAST and Antibiotic Susceptibility Testing in routine. For positive samples containing Gram positive bacteria the results were not convincing. We did not find any correlation between epithelial cells and contamination.

摘要

背景

我们评估了使用 Sysmex UF-5000 进行流式细胞术是否可以通过快速识别培养前的培养阴性和污染样本来改善尿路感染的诊断,从而减少培养平板工作量和反应时间。我们还评估了是否可以使用 Sysmex UF-5000 上的细菌信息标志来减少抗生素药敏谱的反应时间,以区分革兰氏阳性和阴性细菌,然后对阳性尿液样本进行直接抗生素敏感性测试(dAST)。

方法

在培养平板之前,通过流式细胞术对 1000 个尿液样本进行细菌、白细胞和鳞状细胞分析。将不同截止值下流式细胞术分析的结果与培养平板的结果进行比较。我们评估了 100 个尿液样本的 dAST,这些样本通过流式细胞术分析为阳性,包含革兰氏阳性或革兰氏阴性细菌。

结果

使用细菌计数≥100.000/mL 和白细胞计数≥10/μL 的截止值,流式细胞术预测 42.1%的样本无显著生长。我们发现大多数污染样本中含有较少的鳞状细胞。对于包含革兰氏阴性细菌的 52/56 个阳性样本,dAST 与常规检测相同。总体而言,在 560 个测试抗生素组合中有 555 个具有一致性。

结论

流式细胞术为尿路感染的诊断提供了优势。在到达当天筛选阴性尿液样本可减少培养平板工作量,并缩短阴性样本的反应时间。细菌信息标志可准确预测包含革兰氏阴性细菌的阳性样本进行 dAST,从而可以在到达后的第二天报告抗生素药敏谱。对于包含革兰氏阴性细菌的阳性样本,dAST 和常规抗生素药敏试验之间的一致性非常好。对于包含革兰氏阳性细菌的阳性样本,结果并不令人信服。我们没有发现上皮细胞与污染之间存在任何相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3376/8259986/4daeca1752d6/pone.0254064.g001.jpg

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