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

作者：Meza Menchaca Thuluz^*; Williams John; Rodriguez Estrada Rocio B; Garcia Bravo Aracely; Ramos Ligonio Angel; Lopez Monteon Aracely; Zepeda Rossana C

来源：Sensors, 2013, 13(10): 12975-12993.

DOI：10.3390/s131012975

• HPV-18
• microarray
• variants
• LCR

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%26apos;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.

• 出版日期2013-10