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

Masumoto N, Fujii T, Ishikawa M, Mukai M, Saito M, Iwata T, Fukuchi T, Kubushiro K, Tsukazaki K, Nozawa S - Br. J. Cancer (2003)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

ABSTRACT
A fluid-based Papanicolaou test has been established to improve sample collection and preparation. This study was the first large-scale 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=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.

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Detection of PCR products using electrophoresis. PCR products were electrophoresed on a 3% agarose gel. Lane 1, PCR products using genomic DNA with grade 1 degradation as a template; lane 2, PCR products using genomic DNA with grade 2 degradation as a template; lanes 3, 4, and 5, PCR products using genomic DNA with grade 3 degradation as a template. M shows the 100-bp DNA ladder marker. P shows the positive control. N shows the negative control.
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fig2: Detection of PCR products using electrophoresis. PCR products were electrophoresed on a 3% agarose gel. Lane 1, PCR products using genomic DNA with grade 1 degradation as a template; lane 2, PCR products using genomic DNA with grade 2 degradation as a template; lanes 3, 4, and 5, PCR products using genomic DNA with grade 3 degradation as a template. M shows the 100-bp DNA ladder marker. P shows the positive control. N shows the negative control.

Mentions: Estimating the quality of the extracted genomic DNA. Genomic DNA was electrophoresed on a 0.8% agarose gel. The quality of the extracted genomic DNA was classified into three categories. Genomic DNA samples 1, 2, 4, 6, 7, 9, 12, and 13 were classified as exhibiting grade 1 degradation. Genomic DNA samples 3, 5, 8, and 10 were classified as exhibiting grade 2 degradation. Genomic DNA sample 11 was classified as exhibiting grade 3 degradation. M shows the λDNA/HindIII marker.


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

Masumoto N, Fujii T, Ishikawa M, Mukai M, Saito M, Iwata T, Fukuchi T, Kubushiro K, Tsukazaki K, Nozawa S - Br. J. Cancer (2003)

Detection of PCR products using electrophoresis. PCR products were electrophoresed on a 3% agarose gel. Lane 1, PCR products using genomic DNA with grade 1 degradation as a template; lane 2, PCR products using genomic DNA with grade 2 degradation as a template; lanes 3, 4, and 5, PCR products using genomic DNA with grade 3 degradation as a template. M shows the 100-bp DNA ladder marker. P shows the positive control. N shows the negative control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2741123&req=5

fig2: Detection of PCR products using electrophoresis. PCR products were electrophoresed on a 3% agarose gel. Lane 1, PCR products using genomic DNA with grade 1 degradation as a template; lane 2, PCR products using genomic DNA with grade 2 degradation as a template; lanes 3, 4, and 5, PCR products using genomic DNA with grade 3 degradation as a template. M shows the 100-bp DNA ladder marker. P shows the positive control. N shows the negative control.
Mentions: Estimating the quality of the extracted genomic DNA. Genomic DNA was electrophoresed on a 0.8% agarose gel. The quality of the extracted genomic DNA was classified into three categories. Genomic DNA samples 1, 2, 4, 6, 7, 9, 12, and 13 were classified as exhibiting grade 1 degradation. Genomic DNA samples 3, 5, 8, and 10 were classified as exhibiting grade 2 degradation. Genomic DNA sample 11 was classified as exhibiting grade 3 degradation. M shows the λDNA/HindIII marker.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.

ABSTRACT
A fluid-based Papanicolaou test has been established to improve sample collection and preparation. This study was the first large-scale 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=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.

Show MeSH
Related in: MedlinePlus