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Papillomavirus screening in cervical cell samples using dual‐label dot‐blot analysis
Author(s) -
Potter Colin G.,
Cooper Kum,
Stickland Julia E.,
Ramshaw Anna L.,
O' McGee James D.
Publication year - 1993
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1711710108
Subject(s) - dot blot , southern blot , blot , polymerase chain reaction , microbiology and biotechnology , human papillomavirus , northern blot , biology , western blot , virus , dna , virology , gene , gene expression , medicine , genetics
The presence of human papillomavirus (HPV) in cervical cells is closely related to the development of cervical carcinoma. Detection of virus may be by Southern blot, dot blot or the highly sensitive polymerase chain reaction. Whatever method is employed, there are problems of false negatives due to poor clinical samples in which the DNA may be degraded or is absent altogether. Here we describe a new method of dual labelling for dot blots using a 32 P‐labelled probe for HPV and a 35 S‐labelled probe for human actin genes. The samples were counted on a Betaplate™ flat‐bed scintillation counter and the data analysed to separate the activities of the two isotopes. The counts from the actin probe show whether human DNA is present or not and false negatives from this cause may thereby be eliminated. The counts due to HPV when compared with those for actin give a quantitative measure of HPV abundance for the particular sample and this may have clinical relevance.