Use of universal and type-specific primers in the polymerase chain reaction for the detection and typing of genital human papillomaviruses.
By using polymerase chain reaction (PCR), we have developed a system for type-specific as well as universal detection of genital human papillomaviruses (HPVs). Primers and probes for specific detection of HPV-16, -18 and -33 were synthesized from the E7 open reading frame (ORF). They were capable of detecting corresponding HPV types with high specificity and sensitivity. Primers for detection of a broad spectrum of HPV (universal primers) were synthesized from the L1 ORF. The universal primers were shown to be capable of amplifying HPV-6b, -11, -16, -18, -33, -52b and -58. The system was applied to various cervical tissue specimens from Japanese patients. They consisted of 26 normal specimens, 18 from cervical dysplasias and 29 from cervical carcinomas. HPV was detected in none of the normal specimens. On the other hand, many of the specimens from cervical dysplasias and carcinomas were found to be positive for HPV, especially HPV-16. Except for one, all the specimens which were positive with the type-specific PCRs were also positive with the universal PCR. Furthermore, substantial numbers of specimens were found to be positive only with the universal PCR. Cloning and sequencing of DNA segments amplified by the universal primers were undertaken to characterize some of the unknown HPVs. Our PCR system may thus be useful for the specific detection of the three major types of oncogenic HPVs and also for the detection of a broad spectrum of HPVs including possibly novel HPV types.
Maki
H
H
Shionogi Institute for Medical Science, Osaka.
Saito
S
S
Ibaraki
T
T
Ichijo
M
M
Yoshie
O
O
D002460:Cell Line; D003001:Cloning, Molecular; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D002578:Uterine Cervical Dysplasia / Q000235:genetics* / Q000382:microbiology*; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000382:microbiology*