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Fast solid support detection of PCR amplified viral DNA sequences using radioiodinated or hapten labelled primers
S Sauvaigo1, B Fouqué, A Roget
1CIS BIO International, Département de Recherche Fondamentale, Grenoble, France.
Abstract:
Oligonucleotides with novel modifications have been synthesized and incorporated into enzymatically 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 colorimetric 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.