A multiple luminescent procedure for the detection of different papillomaviruses on dot blots.
We developed a method based on the use of various luminescent systems for identification of several nucleic acid sequences on the same dot blots. In a simultaneous assay, the target DNA molecules were hybridized with different DNA probes. These probes were labelled with biotin or digoxigenin or directly coupled to enzymes such as glucose-6-phosphate dehydrogenase, peroxidase or alkaline phosphatase. After hybridization, these labels were detected by luminescent reactions using an amplified camera. Rapid detection and specific identification of pathogenic agents such as human papillomaviruses (HPV) could be performed in a single step by this procedure. Polymerase chain reaction was carried out using general primers and virus types were identified on dot blots using short HPV 11, 16 and 18 specific oligonucleotides.
Chikhaoui
Y
Y
INSERM Unité 58, Montpellier, France.
Balaguer
P
P
Terouanne
B
B
Boussioux
A M
AM
Nicolas
J C
JC
D001483:Base Sequence; D005260:Female; D006241:Haptens; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D027383:Papillomaviridae / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis*
D006241:Haptens; D015345:Oligonucleotide Probes