Detection of human papillomavirus in squamous intraepithelial lesions by consensus and type-specific polymerase chain reaction.
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.
- [show abstract] [hide abstract]
ABSTRACT: Uterine cervix cancer is the second most frequent female cancer after breast cancer in Morocco and represents a public health problem. Cervical cancer is highly linked to human papillomavirus (HPV) especially types 16 and 18 which are the highly oncogenic genotypes. To identify the contribution of HPV testing in the prevention of cervical cancer in Morocco, 147 biopsies collected at the Institut National d'Oncologie and 447 swabs from pathology laboratories and gynaecologist offices in Rabat were HPV analysed. HPV testing were made without any presumption of the histopathological diagnosis. A total of 147 paraffin-embedded biopsies and 447 exfoliated cervical samples were included. Based on histopathology results of the 147 biopsies, most cervical lesions were invasive carcinomas and non specific inflammations (NSI). With the molecular assay, HPV was detected in 91/147 (62%) patients. The high risk types 16 and 18 were found in 45% of the cases (41/91) and HPV 18 in 19% of the cases (17/91). Double infection with HPV 16 and 18 was found in 3 cases. Among the 447 swabs tested, 28 were HPV positives. Cytology results showed that 46% were inflammations (13/28). Among them, 10 patients had a NSI and only 3 patients had a cytology diagnosis of HPV infection. Based on these data, HPV testing should be associated to cervical cytology screening according to two algorithms established in function of the age of the patient and viral natural history. Combination of cytology and HPV testing allow identification of patient with high risk for development of high grade cervical lesions and improve cervical cancer prevention.Journal of Clinical Virology 09/2003; 27(3):286-95. · 3.29 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Chronic rejection represents a major cause of long-term kidney graft loss. T cells that are predominant in long-term rejected kidney allografts (35 ± 10% of area infiltrate) may thus be instrumental in this phenomenom, which is likely to be dependant on the indirect pathway of allorecognition only. We have analyzed the variations in T cell repertoire usage of the Vβ chain at the complementary determining region 3 (CDR3) level in 18 human kidney grafts lost due to chronic rejection. We observed a strongly biased intragraft TCR Vβ usage for the majority of Vβ families and also a very high percentage (55%) of Vβ families exhibiting common and oligoclonal Vβ-Cβ rearrangements in the grafts of patients with chronic rejection associated with superimposed histologically acute lesions. Furthermore, Vβ8 and Vβ23 families exhibited common and oligoclonal Vβ-Jβ rearrangements in 4 of 18 patients (22%). Several CDR3 amino acid sequences were found for the common and oligoclonal Vβ8-Jβ1.4 rearrangement. Quantitative PCR showed that biased Vβ transcripts were also overexpressed in chronically rejected kidneys with superimposed acute lesions. In contrast, T lymphocytes infiltrating rejected allografts with chronic rejection only showed an unaltered Gaussian-type CDR3 length distribution. This pattern suggests that late graft failure associated with histological lesions restricted to Banff-defined chronic rejection does not involve T cell-mediated injury. Thus, our observation suggests that a limited number of determinants stimulates the recipient immune system in long-term allograft failure. The possibility of a local response against viral or parenchymatous cell-derived determinants is discussed.The Journal of Immunology 02/2000; 164(3):1553-1563. · 5.52 Impact Factor
- Journal of Bacteriology and Virology 01/2009; 39(4).