Menstrual Blood Testing With Capture Sequencing Detects High-Risk Human Papillomavirus

Persistent infection of high-risk human papillomavirus (hrHPV) is associated with various HPV-related pre-cancers and cancers. In recent years, HPV testing has gradually become the primary method for cervical cancer screening.

Self-sampling HPV testing is a proposed alternative cervical cancer screening for avoiding stigma and improving participation. Most existing self-sampling HPV studies were based on various sampling brushes inserted into the vagina, and patients may experience discomfort during sampling. Compared with these methods, menstrual blood (MB) collection is associated with less stigma and pain.


Gynecologists at the Sun Yat-sen University (Guangzhou, China) and their colleagues recruited women who were premenopausal and had tested positive for hrHPV in the Central Hospital of Wuhan (Wuhan, China), from September 1, 2020, to April 1, 2021. Cervical smears were collected from recruited patients by an experienced gynecologist. Self-provided commercial sanitary pads were used to collect MB from enrolled patients. A total of 120 enrolled women with hrHPV (mean [SD; range] age, 33.9 [6.9; 20.0 -52.0] years) provided 137 sanitary pads.

A commercial Tiangen dried blood spot DNA extraction kit (Tiangen Biotech, Beijing, China) was used to extract MB DNA from sanitary pads. The concentration of DNA was confirmed using the Qubit 4.0 fluorometer and Qubit dsDNA high-sensitivity (HS) assay kit (Thermo Fisher Scientific, Waltham, MA, USA), and the purity of DNA was determined by Thermo Fisher Scientific’s NanoDrop One spectrophotometer. Extracted MB DNA was stored at −20 °C. Sequencing was performed using a NovaSeq PE150 (Illumina, San Diego, CA, USA). The 21 HPV GenoArray (GA) test (Hybribio Ltd., Hong Kong, China) was used for genotyping.

The investigators reported that the overall concordance rate of MB hrHPV capture sequencing and cervical HPV testing was 92.7, with a κ value of 0.763. Among 24 samples with incomplete concordance or discordant results, 11 samples with additional hrHPV genotypes (45.8%), five true-negative samples (20.8%), and the correct hrHPV genotypes of two samples (8.3%) were correctly identified by MB hrHPV capture sequencing. MB hrHPV detection of hrHPV was equivalent on different menstrual cycle days (MCDs), with an MB hrHPV–positive rate of 27/28 patients (96.4%) for MCD 1, 52/57 patients (91.2%) for MCD 2, 27/28 patients for MCD 3, 4/4 of patients (100%) for MCD 4, and 3/3 patients (100%) for MCD 5. The sensitivity of the MB hrHPV capture sequencing was 97.7%.

The authors concluded that MB hrHPV capture sequencing is a feasible and accurate self-collected approach for cervical cancer screening. This study found that this method is associated with superior performance in identification of HPV genotypes and true-negative events compared with cervical HPV testing. The study was published on December 23, 2021 in the journal JAMA Network Open.

Related Links:
Sun Yat-sen University
Central Hospital of Wuhan
Tiangen Biotech
Thermo Fisher Scientific
Illumina
Hybribio Ltd