The preparation of quality control materials for human papillomavirus genotyping

Abstract

Objective To prepare the HPV genotyping control materials. Methods Three hundred cervical smears with different HPV genotypes were collected and detected by surface plasmon resonance and sequencing. The primers for specific genotype were designed according to GenBank. The recombinant plasmid was constructed through PCR amplification, ligation and transformation. Thirty recombinant plasmids were identified through PCR amplification, sequencing, and the sequences were compared using BLAST. Results The collected HPV infectious samples contained 30 different genotypes including HPV 6,16,18 and so on.The fragment sequences of PCR amplification were concordant with the designed. The fragment sizes of the other types ranged from 1 500 to 2 000 bp except HPV CP8304. And the 30 recombinant plasmids identified by PCR were concordant with the target. The identity of BLAST was 99%. In the fragment of 1500 bp in length, 11 bases were inconsistent with the reference sequence. Conclusions Genotyping control materials were developed successfully. The human papillomavirns genotyping control materials covered all the most common types and included 14 types with high-risk, 3 types with medium-risk and 13 types with low-risk. Key words: Papillomaviridae; Genotype; Reagent kits,diagnostic