优化快速和敏感的核酸侧流生物传感器用于乙型肝炎病毒检测
Abbas Ali Husseini1,2, Serap Yesilkir Baydar3,4
1Life Sciences and Biomedical Engineering Application and Research Centre, Istanbul Gelisim University, Istanbul, Türkiye. husseini.hazara@gmail.com.
Molecular biology reports
|August 17, 2023
概括
这项研究开发了一种快速,具有成本效益的侧流生物传感器,用于在护理点检测乙型肝炎病毒 (HBV). 来自血清的直接放大显示出比纯化核酸更低的检测极限,为资源有限的环境提供了一个有希望的,尽管不那么敏感的解决方案.
科学领域:
- 生物技术是生物技术.
- 分子诊断学 分子诊断学
- 照顾点检测检测 照顾点检测
背景情况:
- 目前正在探索从 lysate 中直接放大核酸的方法,用于点诊断 (POC) 诊断.
- 乙型肝炎病毒 (HBV) 检测测定对疾病管理至关重要.
研究的目的:
- 开发一种快速,精确,经济高效的侧流生物传感器,用于在护理点检测HBV核酸.
- 评估直接放大与净化核酸在循环介导同热放大与侧向流 (LAMP-LF) 生物传感相结合的有效性.
主要方法:
- 使用临床血清样本通过实时PCR确认HBV阳性.
- 核酸是使用旋转柱净化或直接热处理血清来制备的.
- 循环介导的异热放大 (LAMP) 初始剂针对HBV聚合酶基因的250bp片段.
- 确定了最佳的LAMP条件 (67°C30分钟).
- 检测是使用横向流量测定 (Milenia HybriDetect1条纹) 进行的.
主要成果:
- 在67°C30分钟的优化LAMP反应显示出显著的度.
- 使用净化的核酸,侧流检测检测到病毒载荷从108 IU/ml降至101 IU/ml.
- 从经热处理的血清直接放大检测到的最低病毒载量为103 IU/ml.
结论:
- LAMP-LF测定是资源有限的环境中HBV测试的一个有希望的工具.
- 没有DNA净化的直接放大可能会降低测试的灵敏度.
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