In situ amplification of single copy gene segments in individual cells by the polymerase chain reaction.
A method is described using the polymerase chain reaction (PCR) to amplify defined nucleic acid strands in individual cells in situ in conventional smears of bone marrow and peripheral cells. Using radioactively labeled precursors, the incorporation into newly synthesized strands by PCR can be detected by microautoradiography. The specificity of the method can be monitored by gel electrophoresis of the material shed into the reaction mixture. Thus it could be shown that even single genes in individual cells can be amplified to visibility. In a mixture of HIV infected and non infected cells both can be clearly distinguished from one another.
Medizinische Klinik Innenstadt der Ludwig Maximilians Universität, München, Germany.
W
W
Spann
K
K
Pachmann
H
H
Zabnienska
A
A
Pielmeier
B
B
Emmerich
D001345:Autoradiography / Q000379:methods* / Q000592:standards; D004586:Electrophoresis; D006801:Humans; D008852:Microradiography; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity