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一种利用γ-谷氨酰转肽酶激活型荧光探针快速检测幽门螺杆菌感染的新方法。

A novel method for rapid detection of a Helicobacter pylori infection using a γ-glutamyltranspeptidase-activatable fluorescent probe.

机构信息

Department of Gastroenterology and Hepatology, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki, 852-8501, Japan.

Divison of Medicine and Clinical Science, Faculty of Medicine, Tottori University, 36-1 Nishi-cho, Yonago, 683-8504, Japan.

出版信息

Sci Rep. 2019 Jul 1;9(1):9467. doi: 10.1038/s41598-019-45768-x.

DOI:10.1038/s41598-019-45768-x
PMID:31263136
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6603024/
Abstract

A γ-glutamyl hydroxymethyl rhodamine green probe (gGlu-HMRG) reacts with γ-glutamyltranspeptidase (GGT) and immediately produces fluorescence, is clinically applied for real-time cancers' visualization. Since Helicobacter pylori produces GGT, this study aimed to investigate whether gGlu-HMRG can be used to detect H. pylori infections. A wild-type H. pylori strain and the ggt gene-disrupted mutant were cultured and treated with gGlu-HMRG. This fluorescent probe assay was used to quantify GGT activity of H. pylori ex vivo using gastric biopsy specimens. The H. pylori diagnostic capabilities of the assay were determined from altered fluorescence intensity (FI) values at 5 min (FIV-5) and 15 minutes (FIV-15). Distinct fluorescence was identified in wild H. pylori strain, using gGlu-HMRG, whereas no fluorescence was observed in ggt gene-disrupted mutant strain. On ex vivo imaging of gGlu-HMRG, fluorescence intensity increased markedly with time in H. pylori-positive specimens; however, the H. pylori-negative specimens displayed a slight increase in FI. FIV-5 and FIV-15 differed significantly between H. pylori-positive and -negative specimens. FIV-15 differed significantly between H. pylori-positive and -eradicated group. This assay sensitivity and specificity were 75.0% and 83.3% in the antrum and 82.6% and 89.5% in the stomach body. GGT-activatable fluorescence probe is applicable for rapid diagnosis of H. pylori.

摘要

一种 γ-谷氨酰羟甲基罗丹明绿探针 (gGlu-HMRG) 与 γ-谷氨酰转肽酶 (GGT) 反应立即产生荧光，临床上用于实时癌症可视化。由于幽门螺杆菌产生 GGT，本研究旨在探讨 gGlu-HMRG 是否可用于检测 H. pylori 感染。培养野生型 H. pylori 菌株和 ggt 基因缺失突变体，并使用 gGlu-HMRG 处理。使用该荧光探针检测法，通过胃活检标本检测 H. pylori 的 GGT 活性。通过 5 分钟（FIV-5）和 15 分钟（FIV-15）时荧光强度（FI）值的变化（FIV-5 和 FIV-15）来确定检测的 H. pylori 诊断能力。使用 gGlu-HMRG 可在野生 H. pylori 菌株中识别出明显的荧光，而在 ggt 基因缺失突变体菌株中未观察到荧光。在 gGlu-HMRG 的体外成像中，阳性标本的荧光强度随时间显著增加；然而，阴性标本的 FI 略有增加。FIV-5 和 FIV-15 在 H. pylori 阳性和阴性标本之间有显著差异。FIV-15 在 H. pylori 阳性和根除组之间有显著差异。该检测法在胃窦和胃体的敏感性和特异性分别为 75.0%和 83.3%和 82.6%和 89.5%。GGT 激活荧光探针可用于快速诊断 H. pylori。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/359c7496c11f/41598_2019_45768_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/79e8475921f0/41598_2019_45768_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/358c558dbcf1/41598_2019_45768_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/b91bfec5c865/41598_2019_45768_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/4296cc6a35d0/41598_2019_45768_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/3d64ca479ae0/41598_2019_45768_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/5b1d8e04f6e4/41598_2019_45768_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/359c7496c11f/41598_2019_45768_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/79e8475921f0/41598_2019_45768_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/358c558dbcf1/41598_2019_45768_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/b91bfec5c865/41598_2019_45768_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/4296cc6a35d0/41598_2019_45768_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/3d64ca479ae0/41598_2019_45768_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/5b1d8e04f6e4/41598_2019_45768_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1068/6603024/359c7496c11f/41598_2019_45768_Fig7_HTML.jpg

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