Detection of Aeromonas hydrophila possessing aerolysin gene using gold nanoparticle probe.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-12-31 eCollection Date: 2023-12-01 DOI:10.5455/javar.2023.j714

Arren Christian M de Guia, Mary Rose D Uy-de Guia, Juvy J Monserate, Joel R Salazar, Ravelina R Velasco, Claro N Mingala, Karl Marx A Quiazon

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引用次数: 0

Abstract

Objective: The aerolysin (aerA) is a virulence indicator used to identify the pathogenicity of the Aeromonas strain. Targeting a pathogen's crucial virulence gene for detection is essential, as it determines the potential threat to the host. This study aimed to develop a gold nanoparticle (AuNP) probe for detecting the gene aerA in Aeromonas hydrophila among field samples.

Materials and methods: Kidney samples among both healthy and sick Nile tilapias in five provinces of Luzon Island were collected for bacterial analysis. Screening using specific primers targeting aerA was conducted in parallel with testing the AuNPs probe on the same sample set. The positive control provided by BFAR-NFLD, confirmed by polymerase chain reaction (PCR) assay, was used as a positive sample containing the target gene.

Results: The AuNP probe demonstrated a computed accuracy of 81.32%, sensitivity of 100%, and specificity of 81.26%. Among the 257 reactions, 59 were false positives, while no false negative results were observed. The AuNP probe could detect aerA at levels as low as 30 ng/µl. The low prevalence of the target gene may be attributed to the use of general media instead of specific media like Rimler-Shotts agar.

Conclusion: The established colorimetric detection method for A. hydrophila with the aerA gene offers a swift alternative to PCR, negating the requirement for advanced equipment like a thermal cycler.

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利用金纳米粒子探针检测拥有溶气素基因的嗜水气单胞菌。

目的：气溶素（aerA）是一种毒力指标，用于确定气单胞菌株的致病性。针对病原体的关键毒力基因进行检测至关重要，因为它决定了病原体对宿主的潜在威胁。本研究旨在开发一种金纳米粒子（AuNP）探针，用于检测野外样本中嗜水气单胞菌的 aerA 基因：材料：收集吕宋岛五个省健康和患病尼罗罗非鱼的肾脏样本进行细菌分析。在使用针对 aerA 的特异性引物进行筛选的同时，还在同一组样本中测试了 AuNPs 探针。由 BFAR-NFLD 提供并经聚合酶链反应（PCR）测定确认的阳性对照被用作含有目标基因的阳性样本：AuNP 探针的计算准确率为 81.32%，灵敏度为 100%，特异性为 81.26%。在 257 个反应中，59 个为假阳性，没有发现假阴性结果。AuNP 探针可在低至 30 纳克/微升的水平上检测到 aerA。目标基因的低流行率可能是由于使用了普通培养基而不是 Rimler-Shotts 琼脂等特定培养基：已建立的带 aerA 基因的蚜虫比色检测方法提供了一种快速的 PCR 替代方法，无需热循环仪等先进设备。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.