Evaluation of non-radioactive temperature gradient SSCP analysis and of temperature gradient gel electrophoresis for the detection of HPV 6-variants in condylomata acuminata and Buschke-Loewenstein tumours.
A modified non-radioactive single strand conformation polymorphism analysis incorporating a temperature gradient (TG-SSCP) and temperature gradient gel electrophoresis (TGGE) were evaluated for the detection of human papillomavirus type 6 (HPV 6)-variants in 41 condylomata acuminata and 5 Buschke-Loewenstein tumours. TG-SSCP and TGGE analysed part of the transforming ORF E6 of HPV 6 spanning nucleotides 10 to 495. TG-SSCP distinguished between 8 HPV 6-variants whereas TGGE demonstrated 6 different DNA-species. HPV 6-strains found in Buschke-Loewenstein tumours did not vary in the analysed portion of the E6 ORF as compared to ordinary condylomata acuminata. TG-SSCP and TGGE further showed absence of double infection with different HPV 6-strains in the analysed samples. Our results demonstrated that both methods may be successfully used for the detection of different strains of microbiological agents, although TG-SSCP seemed to provide easier execution and to confer a higher degree of flexibility than TGGE.
Rübben
A
A
Department of Dermatology, RWTH-Aachen, Germany.
Traidl
C
C
Baron
J M
JM
Grussendorf-Conen
E I
EI
D003218:Condylomata Acuminata / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005069:Evaluation Studies as Topic; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D009711:Nucleotides / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA; D013696:Temperature; D014412:Tumor Virus Infections / Q000821:virology*
D004279:DNA, Viral; D009711:Nucleotides