Detection of human papillomavirus in squamous intraepithelial lesions by consensus and type-specific polymerase chain reaction.

Centre Hospitalier Universitaire de Nancy, Nancy, Lorraine, France
European Journal of Obstetrics & Gynecology and Reproductive Biology (Impact Factor: 1.63). 01/1994; 52(3):193-200. DOI: 10.1016/0028-2243(93)90071-J
Source: PubMed

ABSTRACT Polymerase chain reaction (PCR) was used to identify human papillomavirus (HPV) in 216 cervical biopsy specimens from women referred to the gynecological out-patient unit for colposcopy because of an abnormal smear. HPV DNA was screened using type-specific primers for HPV6, 11, 16, 18, 31 and 33 (TS-PCR) as well as a consensus primer located in the E1 region of the HPV genome (C-PCR). TS-PCR specificity was validated by Southern blot analysis. Low-grade (SIL 1) and high-grade (SIL 2) squamous intraepithelial lesions were found in 165 biopsies. HPV16 detection was better with PCR than Southern blot, particularly for SIL 1 and SIL 2. The fact that 10% of HPV16 (all SIL 2) were not detected by C-PCR indicates that both PCR techniques should be performed. C-PCR also detects uncharacterized HPV types (8.6% prevalence in our results), mainly in SIL 1 and SIL 2. HPV16, the most frequently isolated type (prevalence 21%), was associated with SIL 2 in 83% of cases. A low HPV prevalence was found in specimens without dysplastic cells. These results suggest that PCR may be an important tool for identifying women at risk for developing dysplasia or cervical cancer.

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    ABSTRACT: The infections by human papillomaviruses (HPVs) are clearly associated with the subsequent development of cervical cancer. In this study, HPV genotype distribution and prevalence were detected in Korean women from January to December 2008 using PCR-DNA sequencing. A total of 2,562 cervical samples from Korean women having routine Pap smear cytology screening were used. HPV DNA was extracted from cervical swab samples and amplified by PCR in L1 region of HPV. HPV DNA was detected in 23.2% and 65.5% from the groups of normal and abnormal Pap cytology, respectively. The prevalence of high-risk types of HPV had the highest frequency in the <30 year-olds' group (50.6%). The prevalence of HPV in normal, ASCUS, LSIL and HSIL groups was 23.2%, 58.1%, 96.3% and 97.0%, respectively. Moreover, the frequencies of the high-risk types of HPV were 16.2% in the normal Pap cytology, 44.7% in the ASCUS, 76.1% in the LSIL and 94.1% in the HSIL groups. The prevalence of the high-risk types of HPV increased in proportion to the severity of the cytological classification. In the HSIL group, HPV type 16 was the most frequently found at 32.4%, followed by types 58, 53 and 33 at 17.6%, 14.7% and 11.8%, respectively. HPV type 82 was found in 5.6% of the HSIL group and was not detected in the normal Pap cytology group. The frequency of high-risk type of HPV 82 is firstly reported in Korean women. This finding could be an informative basis for the development of future HPV vaccination strategies in Korean women.
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    ABSTRACT: Some HPV types are strongly associated with cervical cancer. The HPV DNA typing method is actually a complicated procedure. We developed an original method using a consensus polymerase chain reaction and a colorimetric microtiter plate hybridization assay to automate detection of amplified PCR products. This methodology has a good sensibility and specificity and improve the practicability, reproducibility and rapidity of HPV DNA typing assay.
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    ABSTRACT: Human Papilloma viruses (HPVs) are etiological agents for cervical cancer and are classified into low- and high-risk categories. The aim of this study was to determine the frequency of the HPV genotype in the HPV screening test of Korean women using PCR-direct sequencing. Consensus primers of L1 legion were used for the amplification of HPV DNA and the PCR products (450 bps) obtained were analyzed by automatic sequencing. Sequences were compared with those in GenBank by using the BLAST program. Cervical swab samples of 3,978 women (20-73 years) were tested and the average age was 37.6 years. In this study, 1,174 samples were HPV positive out of 3,978 cervical swab samples screened (29.5%) and 136 samples (11.6%) showed a double infection. A total of 1,310 HPV genotypes were analyzed. The HPV positive rate was the lowest in the 20 years group (69.5%) and most of the samples of the > 60 years group were found HPV positive. Among thirty seven different HPV types identified by sequencing, 21 were HPV high risk types and 16 HPV low risk types were 69.8% (914/1,310) and 26.0% (340/1,310), respectively. In HPV high-risk types, 16 (13.21%), was the most frequently found. HPV 53 (9.62%) and 58 (9.24%) were also frequently found. This group was followed by HPV types 70 (5.50%), 33 (4.73%), 66 (4.20%), 18 (4.05%), 52 (4.05%), 31 (3.97%) and 56 (3.51%) in descending order of frequency. Among HPV low-risk types, 62 (4.20%), 6 (3.59%), 81 (3.59%), 84 (3.51%), and 11 (2.6%) were frequently found. In conclusion, PCR-direct sequencing could be used for quick and reliable typing of known and novel HPVs from clinical specimens. This data could be useful for epidemiological study of HPV and it also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.