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A Low Density Microarray Method for the Identification of Human Papillomavirus Type 18 Variants
Laboratory of Molecular Biology, Health Services Studies Centre, University of Veracruz, 147 Carmen Serdan St., Centre, Veracruz-Llave, Veracruz 91700, Mexico
Biomedical Research Centre, University of Veracruz, Av. Luis Castelazo Ayala St. Xalapa-Enriquez, Veracruz 91120, Mexico
LADISER Inmunology and Molecular Biology, Faculty of Chemical Sciences, University of Veracruz, Orizaba, Veracruz 94340, Mexico
Department of Biochemistry, Biosciences Institute, University College Cork, College Road, Cork, Ireland
* Author to whom correspondence should be addressed.
Received: 9 July 2013; in revised form: 8 August 2013 / Accepted: 30 August 2013 / Published: 26 September 2013
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.