Study On Genotyping Of Virus Related To Cervical Cancer

In women worldwide, cervical cancer is still one of the most common cancers.Infection with human papillomavirus (HPV), particularly HPV16and HPV18, is themain cause of invasive cervical cancer, although other factors such as herpes simplexvirus (HSV) may act in conjunction with HPV. Rapid diagnosis of HPV and HSVlevels has come to be regarded as the most key on the prevention and control andetiological study of cervical cancer. This research based on genotyping of virus relatedcervical cancer, and established a novel multiple fluorescent quantification PCR（FQ-PCR） techniques for the accurate, sensitive and rapid detection of HPV16,HPV18, HSV1and HSV2. In the future, this multiple FQ-PCR assay will assist incervical cancer screening, viral treatment evaluation and epidemiological studies inwhich high throughput analysis is required.To explore the possibility of developing a system for rapid diagnosis and clinicalscreening of cervical cancer, we developed a multiple FQ-PCR assay that cansimultaneously detect and quantify HPV16, HPV18, HSV1and HSV2. To evaluate itspossibilities and practical uses,177samples collected from patients with suspectedHPV and HSV infection in exfoliated cervical cells, genital herpes or labial herpeswere tested by multiple FQ-PCR and compared with results obtained by DNA sequencing. Each virus was detected over a range from1.0×10~1to1.0×10~7copies/reaction. The clinical sensitivity was100%for HPV16, HPV18, HSV1andHSV2. The clinical specificity was97.1%for HPV16,98.1%for HPV18,97.0%forHSV1and96.0%for HSV2. The absolute agreement was97.7%for HPV16,98.3%for HPV18,97.7%for HSV1and97.2%for HSV2; The kappa value was0.96forHPV16,0.92for HPV18,0.94for HSV1and0.93for HSV2, when DNA sequencingwas used as the reference standard. The correlation between the multiple FQ-PCR andsingle FQ-PCR of positive concentration samples was excellent (R~2=0.970forHPV16, R~2=0.965for HPV18, R~2=0.969for HSV1, and R~2=0.966for HSV2).233normal controls and567cases (333of cervicitis,210of cervicalintraepithelial neoplasia (CIN) and24of squamous cell carcinoma (SCC)) in cervicalexfoliative cells were analyzed for HPV16, HPV18, HSV1and HSV2DNA usingthe novel multiple FQ-PCR method. In contrast to normal women, positive rate ofsingle HPV16is related significantly to cervical lesions (odds ratios (ORs)=3.4,p<0.01for cervicitis; ORs=6.2, p<0.01for CIN; ORs=16.9, p<0.01for SCC). positiverate of single HPV18is related significantly to cervical lesions (ORs=2.9, p<0.01forcervicitis; ORs=6.4, p<0.01for CIN; ORs=12.0, p<0.01for SCC). HSV2prevalencein CIN and SCC was significantly higher than in normal women (ORs=2.6, p<0.01for CIN; ORs=2.5, p<0.01for SCC). HSV2coinfection with HPV (HPV16and/orHPV18) in CIN and SCC was strongly higher than in normal women (ORs=7.0,p<0.01for CIN; ORs=30.4, p<0.01for SCC).In summary, we have developed a multiple FQ-PCR assay that has theadvantages of simplicity, rapidity, sensitivity, specificity and high throughputcapability. The assay has potential for developing into a powerful molecular methodfor large scale screening and accurate diagnosis of HPV16, HPV18, HSV1and HSV2in clinical specimens. The presence of HPV is associated closely with cervical cancer,and that HSV2infection or co-infection with HPV might be involved in cervicalcancer development, while HSV1might not be involved.