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Sensors 2013, 13(10), 12975-12993; doi:10.3390/s131012975
Article

A Low Density Microarray Method for the Identification of Human Papillomavirus Type 18 Variants

1,2,3,* , 4
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1 Laboratory of Molecular Biology, Health Services Studies Centre, University of Veracruz, 147 Carmen Serdan St., Centre, Veracruz-Llave, Veracruz 91700, Mexico 2 Biomedical Research Centre, University of Veracruz, Av. Luis Castelazo Ayala St. Xalapa-Enriquez, Veracruz 91120, Mexico 3 LADISER Inmunology and Molecular Biology, Faculty of Chemical Sciences, University of Veracruz, Orizaba, Veracruz 94340, Mexico 4 Department of Biochemistry, Biosciences Institute, University College Cork, College Road, Cork, Ireland
* Author to whom correspondence should be addressed.
Received: 9 July 2013 / Revised: 8 August 2013 / Accepted: 30 August 2013 / Published: 26 September 2013
(This article belongs to the Special Issue Microarray Sensors)
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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 HPV-18; microarray; variants; LCR
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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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.

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