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Nucleic Acids Research, 1990, Vol. 18, No. 11 3175-3183
© 1990

CHEMISTRY

Fast solid support detection of PCR amplified viral DNA sequences using radioiodinated or hapten labelled primers

S. Sauvaigo, B. Fouquè, A. Roget, T. Livache, H. Bazin, C. Chypre and R. Tèoule

Laboratoire des Sondes Molèculaires, CIS BIO International, Dèpartment de Recherche Fondamentale, Laboratoired de Chimie, Centre d'Etudes Nuclèaires de Grenoble 85X 38041 Grenoble Cedex, France

Received March 19, 1990. Accepted May 4, 1990.

Ollgonucleotides with novel modifications have been synthesized and Incorporated Into enzymatlcally amplified DNA sequences. They allow the fast detection of viral DNA sequences after two rounds of amplification. The hybrids formed are Immobilized by affinity on coated tubes and detected by direct beta (32P) or gamma (125I) counting or by colorimetrlc revelation. The effect of a dilution step between the two amplifications Is studied to obtain optimal sensitivity and specificity. This test Is used to detect Human Papillomavirus types 16 and 18 in cells and biopsies and for the specific colorimetric detection of HIV1 in extracted DNA.


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