Laboratory for Chemistry and Physics of Interfaces, Department of Micro-systems Engineering (IMTEK), University of Freiburg, Georges-Köhler-Allee 103, D-79110 Freiburg, Germany.
J Virol Methods. 2010 Jan;163(1):40-8. doi: 10.1016/j.jviromet.2009.07.027. Epub 2009 Aug 5.
Genotyping of the human papilloma virus (HPV) is from a clinical point of view an important diagnostic task as some genotypes play a major role in the development of cervical carcinoma. So far PCR combined with blotting or in situ labelling is known to be the most accurate and sensitive method for detection and genotyping of HPV infection in clinical samples. However, specificity, cost-efficiency and sensitivity are not always satisfactory. A novel DNA biochip is described based on a plastic substrate, onto which small polymer droplets and single-stranded DNA are printed in the form of microarrays. Immobilisation of all compounds on the chip surface is achieved by a short UV-irradiation process, inducing photochemical reactions in the polymer. The chip designed for this study contains 36 probes for determining 12 common, different HPV genotypes. After isolation of the DNA, PCR and biochip read-out, the chip allows for genotyping of the most common virus strains, which, according to current prevalence studies, cover 85-95% of all infections. Following this approach as little as 10 virus copies can be detected within a short exposure time. Even using paraffin-embedded material and 10(4) copies per PCR are sufficient to allow rapid and reliable HPV genotyping.
从临床角度来看，对人类乳头瘤病毒 (HPV) 的基因分型是一项重要的诊断任务，因为某些基因型在宫颈癌的发展中起着重要作用。到目前为止，聚合酶链反应 (PCR) 结合印迹或原位标记已被公认为检测和基因分型临床样本中 HPV 感染的最准确和最敏感的方法。然而，特异性、成本效益和敏感性并不总是令人满意。本文描述了一种基于塑料基质的新型 DNA 生物芯片，在该基质上，以微阵列的形式打印小的聚合物液滴和单链 DNA。通过短的紫外线照射过程，所有化合物在芯片表面的固定化，在聚合物中诱导光化学反应。本研究设计的芯片包含 36 个探针，用于确定 12 种常见的不同 HPV 基因型。在 DNA 的分离、PCR 和生物芯片读出后，该芯片允许对最常见的病毒株进行基因分型，根据目前的流行率研究，这些病毒株涵盖了所有感染的 85-95%。采用这种方法，在短时间的暴露下，只需 10 个病毒拷贝即可检测到。即使使用石蜡包埋材料和每个 PCR 10(4)个拷贝，也足以实现快速可靠的 HPV 基因分型。
