Abstract: We describe a novel microarray based-method for the screening of oncogenic human papillomavirus 18 (HPV-18) molecular variants. Due to the fact that sequencing methodology may underestimate samples containing more than one variant we designed a specific and sensitive stacking DNA hybridization assay. This technology can be used to discriminate between three possible phylogenetic branches of HPV-18. Probes were attached covalently on glass slides and hybridized with single-stranded DNA targets. Prior to hybridization with the probes, the target strands were pre-annealed with the three auxiliary contiguous oligonucleotides flanking the target sequences. Screening HPV-18 positive cell lines and cervical samples were used to evaluate the performance of this HPV DNA microarray. Our results demonstrate that the HPV-18’s variants hybridized specifically to probes, with no detection of unspecific signals. Specific probes successfully reveal detectable point mutations in these variants. The present DNA oligoarray system can be used as a reliable, sensitive and specific method for HPV-18 variant screening. Furthermore, this simple assay allows the use of inexpensive equipment, making it accessible in resource-poor settings.
Keywords: HPV-18; microarray; variants; LCR
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Meza-Menchaca, T.; Williams, J.; Rodríguez-Estrada, R.B.; García-Bravo, A.; Ramos-Ligonio, Á.; López-Monteon, A.; Zepeda, R.C. A Low Density Microarray Method for the Identification of Human Papillomavirus Type 18 Variants. Sensors 2013, 13, 12975-12993.
Meza-Menchaca T, Williams J, Rodríguez-Estrada RB, García-Bravo A, Ramos-Ligonio Á, López-Monteon A, Zepeda RC. A Low Density Microarray Method for the Identification of Human Papillomavirus Type 18 Variants. Sensors. 2013; 13(10):12975-12993.
Meza-Menchaca, Thuluz; Williams, John; Rodríguez-Estrada, Rocío B.; García-Bravo, Aracely; Ramos-Ligonio, Ángel; López-Monteon, Aracely; Zepeda, Rossana C. 2013. "A Low Density Microarray Method for the Identification of Human Papillomavirus Type 18 Variants." Sensors 13, no. 10: 12975-12993.