Hydroxynaphthol blue-based loop-mediated isothermal amplification for the rapid detection of Burkholderia pseudomallei.

IF 1.5 4区医学 Q3 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

Transactions of The Royal Society of Tropical Medicine and Hygiene Pub Date : 2025-05-23 DOI:10.1093/trstmh/traf053

Wiphat Klayut, Panatda Aramrueang, Benjawan Phetsuksiri, Sopa Srisungngam, Watcharee Saisongkorh, Ballang Uppapong, Janisara Rudeeaneksin

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Abstract

Background: Melioidosis is a potentially life-threatening disease caused by Burkholderia pseudomallei. It is endemic in Southeast Asia and northern Australia and is increasingly distributed in other tropical and subtropical regions. Fast and accurate detection of B. pseudomallei is crucial for initiating prompt and effective treatment.

Methods: Loop-mediated isothermal amplification (LAMP) using hydroxynaphthol blue (HNB) for visual detection has been developed and evaluated to detect B. pseudomallei.

Results: The assay amplified B. pseudomallei TTSS1-orf2 at 63°C with a 60-min reaction and no cross-amplification was observed. The detection limit was 1.85×102 fg/µl of B. pseudomallei DNA or 2.08×102 CFU/ml of B. pseudomallei spiked in blood, sputum and urine samples. Clinical evaluation using 191 residual specimens revealed 100% negative agreement in detecting 48 of 49 culture-positive samples. Overall, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 97.96%, 100%, 99.48%, 100% and 99.30%, respectively, while the κ index showed substantial agreement with the culture method (κ=0.99).

Conclusions: This LAMP assay demonstrated good performance, with easy operation and low cost for the rapid detection of B. pseudomallei. It has the potential to be an alternative molecular tool for the early diagnosis of melioidosis in clinical settings.

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基于羟基酚蓝的环介导等温扩增快速检测假马氏伯克氏菌。

背景：类鼻疽是一种由假马利氏伯克氏菌引起的潜在威胁生命的疾病。它是东南亚和澳大利亚北部的地方病，并越来越多地分布在其他热带和亚热带地区。快速准确地检测假假芽胞杆菌对于及时有效地治疗至关重要。方法：利用羟基酚蓝（HNB）进行环介导等温扩增（LAMP）目视检测，并对其检测假芽孢杆菌进行了评价。结果：在63°C条件下，反应60 min，无交叉扩增。血、痰、尿假芽孢杆菌DNA检出限分别为1.85×102 fg/µl和2.08×102 CFU/ml。使用191份剩余标本的临床评估显示，49份培养阳性标本中有48份100%阴性。总体而言，该方法的敏感性、特异性、准确性、阳性预测值和阴性预测值分别为97.96%、100%、99.48%、100%和99.30%，κ指数与培养法基本一致（κ=0.99）。结论：该LAMP检测方法性能好，操作简便，成本低，可快速检测假假芽孢杆菌。它有潜力成为一种替代的分子工具，早期诊断类鼻疽在临床设置。

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

Transactions of The Royal Society of Tropical Medicine and Hygiene 医学-公共卫生、环境卫生与职业卫生

CiteScore

4.00

自引率

9.10%

发文量

115

审稿时长

4-8 weeks

期刊介绍： Transactions of the Royal Society of Tropical Medicine and Hygiene publishes authoritative and impactful original, peer-reviewed articles and reviews on all aspects of tropical medicine.