detection of human papillomaviruses by polymerase chain reaction and ligation reaction on universal microarray检测人类乳头状瘤病毒聚合酶链反应和结扎反应通用的微阵列.pdfVIP

Detection of Human Papillomaviruses by Polymerase Chain Reaction and Ligation Reaction on Universal Microarray 1 1¤ 2,3 4 5 1 Jarmo Ritari *, Jenni Hultman , Rita Fingerroos , Jussi Tarkkanen , Janne Pullat , Lars Paulin , 2,3 1 2,3 Niina Kivi , Petri Auvinen , Eeva Auvinen 1 Institute of Biotechnology, University of Helsinki, Helsinki, Finland, 2 Department of Virology, Helsinki University Hospital Laboratory, Helsinki, Finland, 3 Department of Virology, Haartman Institute, University of Helsinki, Helsinki, Finland, 4 Department of Pathology, Haartman Institute, University of Helsinki, Helsinki, Finland, 5 Functional Genome Analysis, Deutsches Krebsforschungszentrum, Heidelberg, Germany 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 s