Establishment of a rapid paper-based extraction method for human papilloma virus DNA and its preliminary evaluation

Objective To establish a simple and rapid paper-based extraction method for human papilloma virus (HPV) DNA and to evaluate the method. Methods The plasmid containing HPV type 16 gene fragment was employed as the test subjects, and then 6 different paper materials were compared and selected by ultraviolet spectrophotometry and real-time fluorescence quantitative PCR (qPCR). Then, according to the efficiency of nucleic acid extraction and the optimal binding area, a convenient paper strip was designed. Finally, the designed paper strip was applied to extract HPV DNA from 18 clinical cervical exfoliated cell samples (13 positives and 5 negatives), with its performance being evaluated after comparing with the conventional boiling method. Results The qualitative filter paper with a nucleic acid binding zone of 2 mm×10 mm could successfully extract HPV DNA from clinical cervical exfoliated cells. When this test strip was used for DNA extraction, all the 13 high-risk HPV positives were successfully detected out of the 18 clinical samples, which was consistent with the results of the conventional boiling method. In addition, the extraction time of the test strip was less than 1 min for each sample, with a detection limit of 8×102 copies/mL and relative standard deviation of 1.15%. Conclusion Our paper-based rapid extraction for HPV DNA does not require pipetting, heating and centrifugation, and the eluent can be directly used as the PCR template for amplification, which greatly simplifies the extraction process of HPV DNA from clinical cervical exfoliated cell samples.