Variability in beta-globin and HPV DNA amplification by PCR from fixed tissues.
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.
Park
J S
JS
Department of Gynecology and Obstetrics, Johns Hopkins Medical Institutions, Baltimore, Maryland.
Leake
J F
JF
Sharma
B K
BK
Toki
T
T
Kessis
T D
TD
Ambros
R A
RA
Shah
K V
KV
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
D004279:DNA, Viral; D005914:Globins