基于 LAMP 的电化学平台,用于监测与宫颈癌进展风险较高相关的 mRNA 水平 HPV 基因组整合情况

IF 6.8 3区 医学 Q1 VIROLOGY Journal of Medical Virology Pub Date : 2024-10-17 DOI:10.1002/jmv.70008
Nasim Izadi, Johana Strmiskova, Milan Anton, Jitka Hausnerova, Martin Bartosik
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引用次数: 0

摘要

人类乳头瘤病毒(HPV)是一组与各种癌症(尤其是宫颈癌)相关的双链 DNA 病毒。高危类型的 HPV 通过将其 DNA 整合到宿主基因组中来显示其致癌潜力。这种整合事件在很大程度上导致了基因组的不稳定性和恶性肿瘤的发展。然而,传统的检测方法,如免疫组化或基于 PCR 的检测,面临着固有的挑战,因此,人们正在开发替代工具,以加快和简化分析。我们的研究介绍了一种创新的生物传感平台,它将环路介导放大与电化学(EC)分析相结合,用于特异性检测 HPV16 整合。通过靶向 E7 mRNA(HPV 整合中的核心分子)和整合后丢失的 E2 病毒基因转录本等关键元素,我们明确区分了外显子型和整合型 HPV16。我们的 EC 数据证实,在整合了病毒基因组的 HPV16 阳性细胞系中,E7 的表达量较高,而在完全整合了基因组的细胞中,E2 的表达量较低。此外,我们还揭示了患者宫颈组织中不同的表达模式,这些模式与数字液滴 PCR、qRT-PCR 或免疫组化染色密切相关。因此,我们的平台有助于深入了解临床样本中的 HPV 整合情况,促进宫颈癌研究和诊断的进一步发展。
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LAMP-based electrochemical platform for monitoring HPV genome integration at the mRNA level associated with higher risk of cervical cancer progression

Human papillomaviruses (HPVs) represent a diverse group of double-stranded DNA viruses associated with various types of cancers, notably cervical cancer. High-risk types of HPVs exhibit their oncogenic potential through the integration of their DNA into the host genome. This integration event contributes significantly to genomic instability and the progression of malignancy. However, traditional detection methods, such as immunohistochemistry or PCR-based assays, face inherent challenges, and thus alternative tools are being developed to fasten and simplify the analysis. Our study introduces an innovative biosensing platform that combines loop-mediated amplification with electrochemical (EC) analysis for the specific detection of HPV16 integration. By targeting key elements like the E7 mRNA, a central player in HPV integration, and the E2 viral gene transcript lost upon integration, we show clear distinction between episomal and integrated forms of HPV16. Our EC data confirmed higher E7 expression in HPV16-positive cell lines having integrated forms of viral genome, while E2 expression was diminished in cells with fully integrated genomes. Moreover, we revealed distinct expression patterns in cervical tissue of patients, correlating well with digital droplet PCR, qRT-PCR, or immunohistochemical staining. Our platform thus offers insights into HPV integration in clinical samples and facilitates further advancements in cervical cancer research and diagnostics.

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来源期刊
Journal of Medical Virology
Journal of Medical Virology 医学-病毒学
CiteScore
23.20
自引率
2.40%
发文量
777
审稿时长
1 months
期刊介绍: The Journal of Medical Virology focuses on publishing original scientific papers on both basic and applied research related to viruses that affect humans. The journal publishes reports covering a wide range of topics, including the characterization, diagnosis, epidemiology, immunology, and pathogenesis of human virus infections. It also includes studies on virus morphology, genetics, replication, and interactions with host cells. The intended readership of the journal includes virologists, microbiologists, immunologists, infectious disease specialists, diagnostic laboratory technologists, epidemiologists, hematologists, and cell biologists. The Journal of Medical Virology is indexed and abstracted in various databases, including Abstracts in Anthropology (Sage), CABI, AgBiotech News & Information, National Agricultural Library, Biological Abstracts, Embase, Global Health, Web of Science, Veterinary Bulletin, and others.
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