Detection of human papillomaviruses by polymerase chain reaction and ligation reaction on universal microarray.

Sensitive and specific detection of human papillomaviruses (HPV) in cervical samples is a useful tool for the early diagnosis of epithelial neoplasia and anogenital lesions. Recent studies support the feasibility of HPV DNA testing instead of cytology (Pap smear) as a primary test in population screening for cervical cancer. This is likely to be an option in the near future in many countries, and it would increase the efficiency of screening for cervical abnormalities. We present here a microarray test for the detection and typing of 15 most important high-risk HPV types and two low risk types. The method is based on type specific multiplex PCR amplification of the L1 viral genomic region followed by ligation detection reaction where two specific ssDNA probes, one containing a fluorescent label and the other a flanking ZipCode sequence, are joined by enzymatic ligation in the presence of the correct HPV PCR product. Human beta-globin is amplified in the same reaction to control for sample quality and adequacy. The genotyping capacity of our approach was evaluated against Linear Array test using cervical samples collected in transport medium. Altogether 14 out of 15 valid samples (93%) gave concordant results between our test and Linear Array. One sample was HPV56 positive in our test and high-risk positive in Hybrid Capture 2 but remained negative in Linear Array. The preliminary results suggest that our test has accurate multiple HPV genotyping capability with the additional advantages of generic detection format, and potential for high-throughput screening.

High prevalence of HPV among female students in Finland.

We studied the prevalence of HPV DNA among university students within the Helsinki metropolitan area in Finland. First-y students attending a general health examination as well as students visiting a general practitioner for contraception were enrolled. Vaginal self-samples or cervicovaginal swabs were collected from a total of 1469 students. HPV DNA was determined using a liquid hybridization test. Of all students 33.0% were positive for HPV DNA. Of all positive samples 84.3% were positive for high-risk HPV. The study demonstrated a strikingly high HPV DNA prevalence among the young women. Self-collected samples proved to be acceptable for HPV DNA detection using liquid hybridization.