title: Variability in beta-globin and HPV DNA amplification by PCR from fixed tissues.
abstract: Paraffin-embedded tissues from a variety of sources and treated with different fixatives were tested for beta-globin and HPV amplification by polymerase chain reaction (PCR). In tests of tissues collected in the previous 2 to 3 yr, excellent rates (87% to 93%) for beta-globin amplification were obtained for specimens fixed in buffered formalin, Bouin's, and Hartmann's solutions. In contrast, the rate of beta-globin amplification was low for tissues fixed in Hollande's solution (7%) and in Hartmann's solution with eosin (33%). The results of beta-globin amplification from archival tissues stored for variable time periods showed no decrease in the amplification rate with longer periods of storage. Human papillomaviruses (HPVs) were identified in 17% of globin negative and in 43% of globin positive tissues. HPV-16 amplification was more efficient when the targeted DNA sequence was small. Variability in amplification depends not only on the type of fixative used, but also on other ill-defined factors. Therefore, conditions for optimal amplification should be determined before undertaking studies of archival material.
authors.affiliation: Department of Gynecology and Obstetrics, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: J S
authors.initials: JS
authors.lastname: Park
authors.forename: J F
authors.initials: JF
authors.lastname: Leake
authors.forename: B K
authors.initials: BK
authors.lastname: Sharma
authors.forename: T
authors.initials: T
authors.lastname: Toki
authors.forename: T D
authors.initials: TD
authors.lastname: Kessis
authors.forename: R A
authors.initials: RA
authors.lastname: Ambros
authors.forename: K V
authors.initials: KV
authors.lastname: Shah
mesh_terms: D015139:Blotting, Southern; D002584:Cervix Uteri / Q000737:chemistry / Q000382:microbiology; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D005260:Female; D005784:Gene Amplification / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics; D005914:Globins / Q000032:analysis / Q000235:genetics*; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D014844:Vulva / Q000737:chemistry / Q000382:microbiology
chemical_list: D004279:DNA, Viral; D005914:Globins