title: Detection of human papillomaviruses by polymerase chain reaction and ligation reaction on universal microarray.
abstract: 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.
authors.affiliation: Institute of Biotechnology, University of Helsinki, Helsinki, Finland. jarmo.ritari@helsinki.fi
authors.forename: Jarmo
authors.initials: J
authors.lastname: Ritari
authors.forename: Jenni
authors.initials: J
authors.lastname: Hultman
authors.forename: Rita
authors.initials: R
authors.lastname: Fingerroos
authors.forename: Jussi
authors.initials: J
authors.lastname: Tarkkanen
authors.forename: Janne
authors.initials: J
authors.lastname: Pullat
authors.forename: Lars
authors.initials: L
authors.lastname: Paulin
authors.forename: Niina
authors.initials: N
authors.lastname: Kivi
authors.forename: Petri
authors.initials: P
authors.lastname: Auvinen
authors.forename: Eeva
authors.initials: E
authors.lastname: Auvinen
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D002583:Uterine Cervical Neoplasms / Q000821:virology*
chemical_list: D017931:DNA Primers