Preparation of long oligonucleotide microarray for detection and sub-typing of human papillomavirus
- VernacularTitle:人乳头瘤病毒分型长链寡核苷酸芯片的制备
- Author:
Min WEI
;
Wenli MA
;
Bao ZHANG
- Publication Type:Journal Article
- Keywords:
papillomavirus, human;
oligonucleotide array sequence;
detection
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To design a microarray of ~60mer oligonucleotide for detection and sub-typing of human papillomavirus (HPV). Methods The type-specific oligonucleotide probes of 4 different types of HPV (6, 11, 16, 18) were designed by using biological software Arraydesigner 2.0, which analyzed the whole genome sequences of HPV and selected optimal probes with high specificity, identical length and similar melting temperature (Tm). These probes were synthesized and printed onto the surface of glass slides in order to prepare a low-density microarray. HPV samples were labeled with fluorescence dyes Cy3 using a method of restriction display PCR (RD-PCR). HPV plasmid DNA was restricted with Sau3AⅠ to produce multiple fragments which were ligated to adaptors subsequently and used as PCR template. PCR labeling was performed with the fluorescently labeled universal primer (Cy3-UP) whose sequence is designed according to the adaptor of the RD-PCR approaches. The labeled samples were then hybridized with the oligonucleotide microarray. Results Both single and multiple HPV DNA samples could be detected with oligonucleotides microarray, and the corresponding HPV subtypes were recognized as well. And no signals were detected in all the negative and blank control spots. Conclusion 60mer oligonucleotide microarray designed by appropriate bioinformatics software can be applied to HPV detection and genotyping on gene level.
