Easy and fast detection and genotyping of high-risk human papillomavirus by dedicated DNA microarrays
Easy and fast detection and genotyping of high-risk human papillomavirus by dedicated DNA microarrays
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DOI：
10.1016/j.jviromet.2006.06.023
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发表时间：
2006-11-01
影响因子：
3.1
通讯作者：
Giordanengo, Valerie
中科院分区：
文献类型：
--
作者：
Albrecht, Valerie;Chevallier, Anne;Giordanengo, Valerie
持续的宫颈高危人乳头瘤病毒（HPV）感染与发生高级别宫颈上皮内病变的风险增加相关。开发了一种两步法用于高危 HPV 的检测和基因分型。首先在 Cy3 标记引物和 dUTP 存在下通过不对称 PCR 扩增 DNA。然后使用 DNA 微阵列杂交对标记的 DNA 进行基因分型。目前的研究评估了实验室设计的 HPV DNA 微阵列对 57 份恶性和非恶性宫颈涂片进行高危 HPV 基因分型的技术功效。该方法针对多种细胞学样本进行了评估：高级鳞状上皮内病变 (HSIL)、低级鳞状上皮内病变 (LSIL) 和高级非典型鳞状细胞 (ASC-H)。在 6 个意义未定的非典型鳞状细胞 (ASC-US) 样本中也检测到了高危 HPV；其中只有一份宫颈标本未受感染，且无组织学病变。 HPV寡核苷酸DNA微阵列基因分型检测到36例单一高危HPV类型感染和5例多种高危类型多重感染。总而言之，这些结果证明了 HPV DNA 微阵列方法的敏感性和特异性。这种方法可以改善宫颈细胞学异常患者的临床管理。 (c) 2006 Elsevier B.V. 保留所有权利。
Persistent cervical high-risk human papillomavirus (HPV) infection is correlated with an increased risk of developing a high-grade cervical intraepithelial lesion. A two-step method was developed for detection and genotyping of high-risk HPV. DNA was firstly amplified by asymmetrical PCR in the presence of Cy3-labelled primers and dUTP. Labelled DNA was then genotyped using DNA microarray hybridization. The current study evaluated the technical efficacy of laboratory-designed HPV DNA microarrays for high-risk HPV genotyping on 57 malignant and nonmalignant cervical smears. The approach was evaluated for a broad range of cytological samples: high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL) and atypical squamous cells of high-grade (ASC-H). High-risk HPV was also detected in six atypical squamous cells of undetermined significance (ASC-US) samples; among them only one cervical specimen was found uninfected, associated with no histological lesion. The HPV oligonucleotide DNA microarray genotyping detected 36 infections with a single high-risk HPV type and 5 multiple infections with several high-risk types. Taken together, these results demonstrate the sensitivity and specificity of the HPV DNA microarray approach. This approach could improve clinical management of patients with cervical cytological abnormalities. (c) 2006 Elsevier B.V. All rights reserved.