Papanicolaou tests and molecular analyses using new fluid-based specimen collection technology in 3000 Japanese women

Abstract

A fluid-based Papanicolaou test has been established to improve sample collection and preparation. This study was the first largescale investigation in Japan to examine the feasibility of using fluid-based Papanicolaou specimens to detect human papillomavirus (HPV) using Hybrid Capture II and polymerase chain reaction (PCR). Three thousand patients who visited Keio University Hospital between October 2000 and February 2001 were enrolled in the study. The results of the fluid-based Papanicolaou tests corresponded well with those of conventional Papanicolaou smears (96.8% concordance). The sensitivities of cervical neoplasia detection using the fluid-based Papanicolaou test (73.9%) and Hybrid Capture II (76.3%, P ¼ 0.55) were not significantly different. Among the cervical intraepithelial neoplasia 3 and squamous cell carcinoma specimens, HPV 16 and HPV 52 were predominantly detected using the PCR method. Although some DNA samples extracted from the fluid-based specimens were degradaded, PCR and direct sequencing could be performed without difficulty even after 1 year of specimen storage. We conclude that fluid-based Papanicolaou specimens can be applied to investigate HPV infection. The fluid-based Papanicolaou specimen collection method is widely used for primary cervical cancer screening (Awen et al, 1994;Roberts et al, 1997;Bolick and Hellman, 1998), largely because of the improvement in specimen quality, the advantage of increased sensitivity, and a reduction in the false negative rate for squamous intraepithelial lesions (Linder and Zahniser, 1997;Park et al, 2001), compared to the conventional Papanicolaou smear method. Fluid-based specimens can be stored at ambient temperature for a longer period; therefore, this collection system offers the advantage that once the specimen has been collected, there is no need to collect additional specimens from the patient to perform a second Papanicolaou test or conduct further investigations. Studies evaluating the clinical utility of Hybrid Capture II (HC II) human papillomavirus (HPV)-DNA testing have also been performed (Clavel et al, 1998;Lin CT et al, 2000a;Yamazaki et al, 2001;Castle et al, 2002). Genital HPV has been reported to be related to cervical cancer carcinogenesis and some types of HPV; HPV 16, for example, is associated with a high risk of cervical neoplasia (Josefsson et al, 2000;Woodman et al, 2001). HC II is a HPV detection test designed to detect 18 types of HPV using microtitre plates and is an appropriate method for HPV screening. We performed the HC II test using fluid-based specimens from patients in whom biopsy studies were also performed to determine whether this method is appropriate for detecting cervical neoplasia in Japan. We also performed HPV typing using fluid-based specimens. We previously reported an HPV-DNA transcript detection method using cytologic specimens and reverse transcriptase-nested polymerase chain reactions (PCR) (Fujii et al, 1995), and a method for detecting multiplex HPV infection using PCR single-stranded DNA-conform...

Do we need a different strategy for HPV screening and vaccination in East Asia?

“…[7][8][9][10][11][12][13][14][15][16][17][18][19][20] For some articles, additional typespecific data were obtained from the authors. 11,12,20 HPV type prevalence data were collected separately for squamous cell carcinoma (SCC) and for adeno-and adenosquamous carcinoma (ADC). Where histological data were not reported, ICC cases were classified as unspecified carcinoma (UC).…”

Ancillary testing of liquid-based cytology specimens for identification of patients at high risk of cervical cancer

“…The HPV genotype was determined by L1-PCR restriction fragment length polymorphism analysis. In patients whose genotype could not be determined by this method, the HPV genotyping was determined by the direct sequencing method [43].…”

Intratumor injection of small interfering RNA-targeting human papillomavirus 18 E6 and E7 successfully inhibits the growth of cervical cancer

“…SKG-II cells were injected into back of the nude mice, and 1 week later visible tumors had developed at the injection sites (tumor volume, 50-60 mm 3 ). To determine the therapeutic effectiveness of E6 and E7 siRNA, intratumoral injection of a mixture of #1 siRNA and atelocollagen or a mixture of GL-2 siRNA and atelocollagen was administered and repeated in 10 days because mixture with atelocollagen and siRNA was stable for this period (15).…”

“…Since human papillomavirus (HPV) infects the uterine cervix and is widely recognized as a risk factor for cervical cancer (2), molecule-targeting therapy of cervical cancer with siRNA targeted against HPV genomes is viewed with hope. HPV genomes have been isolated from >90% of squamous cell carcinomas of the cervix, and HPV16 has been isolated from approximately half of them (3,4). HPV18 is reported to be present in squamous cell carcinoma of the cervix (5) and is one of the most frequent types of HPV isolated from adenocarcinoma of the cervix (6), and is also very frequently isolated from small cell carcinoma (7).…”

“…A total of 2x10 6 SKG-II cells in 0.5 ml of PBS were inoculated with a 26G needle into the back of 8-week-old athymic nude mice obtained from Nihon Clea Japan Inc. One week later, when the tumors had reached an average volume of 50-60 mm 3 , the tumors-bearing mice were treated with #1 siRNA, target for E6 and E7 mRNA, with atelocollagen (Koken Co. Ltd., Tokyo, Japan). The final concentration of atelocollagen was 0.9% and the amount of siRNA was one nanomole.…”