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整合便携式聚合酶链反应和侧向流动分析法用于快速检测16型人乳头瘤病毒

Integration of Portable PCR and a Lateral Flow Assay for the Rapid Detection of HPV Type 16.

作者信息

Yusuf Muhammad, Rohmatulloh Fauzian Giansyah, Soidah Siti, Gaffar Shabarni, Mantilidewi Kemala Isnainiasih, Winarno Gatot Nyarumenteng Adhipurnawan, Hidayat Yudi Mulyana, Subroto Toto

机构信息

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Sumedang, West Java, 45363, Indonesia.

Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, Bandung, West Java, 40133, Indonesia.

出版信息

Med Devices (Auckl). 2025 Jun 26;18:353-360. doi: 10.2147/MDER.S508723. eCollection 2025.

Abstract

BACKGROUND

Human papillomavirus (HPV) is a leading cause of cervical cancer worldwide, with HPV type 16 accounting for roughly half of cases. Although polymerase chain reaction (PCR) is considered the gold standard for HPV detection, its reliance on gel electrophoresis can be costly and require specialized facilities.

OBJECTIVE

We developed a diagnostic approach integrating PCR with a lateral flow assay to detect HPV type 16 using gold nanoparticles as visible labels.

METHODS

Primers targeting the L1 gene of HPV 16 were labeled with 6FAM (forward) and biotin (reverse). Amplified products were applied to lateral flow strips preloaded with streptavidin-gold nanoparticle conjugates. Visible red bands on the test and control lines indicated successful detection.

RESULTS

The optimized assay produced a clear band at ~333 bp by gel electrophoresis and yielded distinct red lines on the lateral flow strip for positive samples. Compared to electrophoresis, this format has a faster turnaround and can reduce costs by eliminating bulky equipment.

CONCLUSION

This simplified and cost-effective method provides a user-friendly alternative to traditional electrophoresis, making it suitable for resource-limited settings. Further large-scale clinical validation is warranted, including cost analyses and multiplexing for additional genotypes.

摘要

背景

人乳头瘤病毒(HPV)是全球宫颈癌的主要病因,其中16型HPV约占病例的一半。尽管聚合酶链反应(PCR)被认为是HPV检测的金标准,但其对凝胶电泳的依赖可能成本高昂且需要专门的设备。

目的

我们开发了一种将PCR与侧向流动分析相结合的诊断方法,以使用金纳米颗粒作为可见标记物来检测16型HPV。

方法

靶向HPV 16型L1基因的引物用6FAM(正向)和生物素(反向)标记。将扩增产物应用于预先加载有链霉亲和素 - 金纳米颗粒偶联物的侧向流动条带。测试线和对照线上可见的红色条带表明检测成功。

结果

优化后的检测方法通过凝胶电泳在约333 bp处产生清晰条带,并且在侧向流动条带上为阳性样品产生明显的红线。与电泳相比,这种形式周转更快,并且通过省去笨重的设备可以降低成本。

结论

这种简化且经济高效的方法为传统电泳提供了一种用户友好的替代方案,使其适用于资源有限的环境。有必要进行进一步的大规模临床验证,包括成本分析和针对其他基因型的多重检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c90a/12208118/b30e26c473e5/MDER-18-353-g0001.jpg

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