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培养过滤蛋白 10 特异性穿孔素在活动性肺结核患儿中的诊断性能。

Diagnostic performance of culture filtered protein 10-specific perforin in pediatric patients with active tuberculosis.

机构信息

Department of Laboratory Medicine, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Department of Laboratory Medicine, Hubei University of Chinese Medicine, Wuhan, China.

出版信息

J Clin Lab Anal. 2020 Nov;34(11):e23477. doi: 10.1002/jcla.23477. Epub 2020 Jul 16.

Abstract

BACKGROUND

Mycobacterium tuberculosis (Mtb)-specific perforin were significantly increased in patients with tuberculosis. This study aims to evaluate the diagnosis value of Mtb-specific perforin in pediatric patients with tuberculosis.

METHODS

Diagnostic performance of perforin levels induced by 6-kDa early secreted antigen target (ESAT6) or culture filtered protein 10 (CFP10) were evaluated in eighty-six samples from children participants by receiver operating characteristic curve analysis. Flow cytometry was used to detect the expression of perforin and INF-γ of CD4 , CD8 T cells in response to CFP10 stimulation.

RESULTS

After ex vivo stimulation, levels of ESAT6/CFP10-specific perforin in LTBI patients were significantly higher than active TB (ATB) patients, non-tuberculosis infection (non-TB), and health control (HC) individuals. The diagnostic efficacy of CFP10-specific perforin for TB diagnosis was significantly higher than ESAT6-specific perforin and T-SPOT assay, and when 0.74 ng/mL was taken as the cutoff value, the sensitivity, specificity, and accuracy were 97.83%, 87.5%, and 93.02%. CFP10-specific perforin in both CD4 and CD8 T cells were significantly higher in ATB patients compared to HCs and further increased in LTBI patients. However, INF-γ was mainly secreted by CD4 T cells and showed no significant difference between LTBI and ATB patients. In addition, CFP10-specific perforin can effectively distinguish between ATB and LTBI with the cutoff value of 1.80 ng/mL. Sensitivity and specificity were 88.46% and 85.62%, respectively.

CONCLUSIONS

CFP10-specific perforin may be used as a novel cellular immunity-based diagnostic marker of pediatric patients with tuberculosis, and with the potential for discriminating ATB from LTBI.

摘要

背景

结核分枝杆菌(Mtb)特异性穿孔素在结核病患者中显著增加。本研究旨在评估 Mtb 特异性穿孔素在儿童结核病患者中的诊断价值。

方法

通过受试者工作特征曲线分析,评估 86 例患儿样本中 6kDa 早期分泌抗原靶标(ESAT6)或培养过滤蛋白 10(CFP10)诱导的穿孔素水平的诊断性能。流式细胞术用于检测 CF10 刺激后 CD4、CD8 T 细胞中穿孔素和 INF-γ的表达。

结果

在体外刺激后,LTBI 患者的 ESAT6/CFP10 特异性穿孔素水平明显高于 ATB 患者、非结核感染(non-TB)和健康对照(HC)个体。CFP10 特异性穿孔素对结核病的诊断效能明显高于 ESAT6 特异性穿孔素和 T-SPOT 检测,当以 0.74ng/mL 为截值时,其灵敏度、特异性和准确性分别为 97.83%、87.5%和 93.02%。与 HC 相比,ATB 患者的 CD4 和 CD8 T 细胞中的 CFP10 特异性穿孔素均显著升高,而 LTBI 患者的穿孔素进一步升高。然而,INF-γ主要由 CD4 T 细胞分泌,LTBI 和 ATB 患者之间无显著差异。此外,CFP10 特异性穿孔素可有效区分 ATB 和 LTBI,截值为 1.80ng/mL,灵敏度和特异性分别为 88.46%和 85.62%。

结论

CFP10 特异性穿孔素可作为儿童结核病患者新型细胞免疫诊断标志物,具有区分 ATB 和 LTBI 的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10e8/7676199/5043ce81c32c/JCLA-34-e23477-g001.jpg

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