A rapid solution hybridization method for detection of human papillomaviruses.
Nucleic acid hybridization methods in routine diagnosis of micro-organisms have been limited by the tedious assay procedures. We have previously described the sandwich hybridization method which allows convenient testing of biological specimens. In this paper we describe the adaptation of the solution hydridization method into the microtitre plate format using 35S-isotope as label. Using 3-hour hybridization followed by 2-hour collection of the hybrids a sensitivity of 5 x 10(5) target DNA molecules was achieved. The method was applied for identification of human papillomaviruses in crude gynaecological specimens. A simple 1-day assay protocol was achieved with high HPV type specificity. The specificity was confirmed by testing a variety of unrelated micro-organisms, none of which gave a positive signal in the test. Results, obtained as numerical values, were easy to interpret; positive and negative samples gave clearly distinguishable signals.
Jalava
T
T
Orion Corporation, Orion Pharmaceutica, Helsinki, Finland.
Kallio
A
A
Leinonen
A W
AW
Ranki
M
M
D002584:Cervix Uteri / Q000382:microbiology; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000302:isolation & purification; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000145:classification / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000382:microbiology; D014626:Vaginal Smears
D015344:DNA Probes, HPV; D004279:DNA, Viral