Biomaterial Database

Simultaneous polymerase chain reaction detection and restriction typing for the diagnosis of human genital papillomavirus infection. 1995

S Pizzighella, and G Pisoni, and F Bevilacqua, and A Vaona, and G Palù

University of Padua, School of Medicine, Microbiology Department, Italy.

A polymerase chain reaction method has been developed which allows the simultaneous detection of the majority of clinically relevant HPV types. Degenerate HPV-specific primers direct the one-step amplification of a DNA region spanning E1 and E7 genes. This enables an immediate distinction between the two groups of papillomaviruses, characterized by high or low oncogenic potential, simply from the size of amplified DNA. The PCR product can be subjected to a second round of amplification with internal primers, which are specific for 7 high-risk HPV types, HPV-16, -18, -31, -33, -35, -45 and -58. Precise identification of one-step or two-step amplified DNA is done by endonuclease digestion with one or two enzymes. The detection sensitivity, which has been assessed using cloned HPV genomes and HeLa and CaSki cell lines, varies from a few tens to a few hundreds of viral genome equivalents. The accuracy of the method has been confirmed by examining cervical scrapings of 44 patients.

UI	MeSH Term	Description	Entries
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D009856	Oncogene Proteins, Viral	Products of viral oncogenes, most commonly retroviral oncogenes. They usually have transforming and often protein kinase activities.	Viral Oncogene Proteins,Viral Transforming Proteins,v-onc Proteins,Transforming Proteins, Viral,v onc Proteins
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D004279	DNA, Viral	Deoxyribonucleic acid that makes up the genetic material of viruses.	Viral DNA
D005260	Female		Females
D005831	Genital Diseases, Female	Pathological processes involving the female reproductive tract (GENITALIA, FEMALE).	Gynecologic Diseases,Female Genital Diseases,Diseases, Female Genital,Diseases, Gynecologic,Female Genital Disease,Genital Disease, Female,Gynecologic Disease
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA