Detection of human papillomavirus type 16 E6/E7 transcripts in fixed paraffin-embedded cervical cancers by the polymerase chain reaction.
A simple, and sensitive method for the detection of human papillomavirus (HPV) 16 E6/E7 transcripts in RNA purified from formalin acetic alcohol (FAA)-fixed, paraffin embedded cervical cancer (CaCx) tissue is described. The entire procedure, including polymerase chain reaction (PCR) amplification of cDNA obtained from mRNA by reverse transcription, can be completed within a day. The results obtained compare favourably with those using total nucleic acids extracted from the same fixed material, and are consistent with results using RNA purified from corresponding fresh tumour tissue. Although developed for investigating the expression of HPV in tumours, this method should find general application in studies of gene expression and viral detection.
Rose
B R
BR
Department of Infectious Diseases, University of Sydney, New South Wales, Australia.
Jiang
X M
XM
Thompson
C H
CH
Tattersall
M H
MH
Cossart
Y E
YE
D004277:DNA, Single-Stranded; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012322:RNA Precursors / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification; D014412:Tumor Virus Infections / Q000235:genetics / Q000382:microbiology*; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000382:microbiology*
D004277:DNA, Single-Stranded; D012322:RNA Precursors; D012367:RNA, Viral