Evaluation of the Roche MagNA Pure 96 nucleic acid extraction platform for the SPF10 line probe assay system

J Appl Microbiol. 2023 Aug 1;134(8):lxad151. doi: 10.1093/jambio/lxad151.

Abstract

Objective: To evaluate the MagNA Pure 96 (MP96) nucleic acid extraction system as an alternative for cervical specimen processing for human papillomavirus (HPV) genotyping detection by reverse hybridization line probe assay (LiPA-25), compared to the well-established extraction system MagNA Pure LC 2.0 (MPLC).

Methods and results: A total of 200 cervical samples preserved in ThinPrepCyt® solution were extracted by MP96 and MPLC, respectively, and then purified nucleic acids were amplified using the SPF10 PCR primer set. Amplification products were subjected to SPF10-DNA enzyme immunoassay (DEIA) and LiPA-25. The concordance between different extraction methods in this study was reflected in the comparison of the results of the DEIA and LiPA-25. Agreement of HPV-positive results (DEIA) was 97.5% (Kappa = 0.932). Pair-wise analyses of either HPV grouping (any HPV genotypes, any high-risk HPV genotypes, any low-risk HPV genotypes, any nonavalent vaccine-targeted HPV, and any non-vaccine-targeted oncogenic HPV genotypes) identified >95% agreement (all Kappa > 0.900). For the two extraction methods, there was no statistical difference (chi-square test: P = 0.690) for single versus multiple genotypes, and concordant, compatible, and discordant genotypes were observed in 87.0%, 9.5%, and 3.5% of 200 samples, respectively.

Conclusion: HPV genotyping results of the MPLC system and the MP96 system showed a high degree of concordance. Combined with the advantages of high-throughput and anti-contamination of MP96, the MP96 extraction system could be more suitable for the testing of samples in future studies.

Keywords: DNA extraction; MagNA Pure 96 System; cervical cancer; human papillomavirus.

MeSH terms

  • Cervix Uteri / chemistry
  • DNA, Viral / genetics
  • Female
  • Genotype
  • Humans
  • Papillomaviridae / genetics
  • Papillomavirus Infections*
  • Polymerase Chain Reaction / methods

Substances

  • DNA, Viral

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