A simple and efficient method for potential point-of-care diagnosis of human papillomavirus genotypes: combination of isothermal recombinase polymerase amplification with lateral flow dipstick and reverse dot blot

Anal Bioanal Chem. 2019 Nov;411(28):7451-7460. doi: 10.1007/s00216-019-02113-5. Epub 2019 Oct 6.

Abstract

Cervical cancer is the second most common cancer in the world's woman population with a high incidence in developing countries where diagnostic conditions for the cancer are poor. The main culprit causing the cancer is the human papillomavirus (HPV). HPV is divided into three major groups, i.e., high-risk (HR) group, probable high-risk (pHR) group, and low-risk (LR) group according to their potential of causing cervical cancer. Therefore, developing a sensitive, reliable, and cost-effective point-of-care diagnostic method for the virus genotypes in developing countries even worldwide is of high importance for the cancer prevention and control strategies. Here we present a combined method of isothermal recombinase polymerase amplification (RPA), lateral flow dipstick (LFD), and reverse dot blot (RDB), in quick point-of-care identification of HPV genotypes. The combined method is highly specific to HPV when the conserved L1 genes are used as targeted genes for amplification. The method can be used in identification of HPV genotypes at point-of-care within 1 h with a sensitivity of low to 100 fg of the virus genomic DNA. We have demonstrated that it is an excellent diagnostic point-of-care assay in monitoring the disease without time-consuming and expensive procedures and devices.

Keywords: Human papillomavirus (HPV); Lateral flow dipstick (LFD); Point-of-care testing (POCT); Recombinase polymerase amplification (RPA); Reverse dot blot (RDB); Visual detection.

MeSH terms

  • Amino Acid Sequence
  • Blotting, Southern / methods*
  • DNA, Viral / analysis
  • DNA, Viral / standards
  • Genes, Viral*
  • Humans
  • Limit of Detection
  • Papillomaviridae / genetics
  • Papillomaviridae / isolation & purification*
  • Papillomavirus Infections / diagnosis*
  • Papillomavirus Infections / virology
  • Point-of-Care Systems*
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results

Substances

  • DNA, Viral