Uniplex E6/E7 PCR method detecting E6 or E7 genes in 39 human papillomavirus types

Okodo, Mitsuaki; Okayama, Kaori; Teruya, Koji; Sasagawa, Toshiyuki

doi:10.1002/jmv.25017

Cited by 31 publications

(61 citation statements)

References 23 publications

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“…The strength of the study was the use of samples in which various HPV genotypes were comprehensively detected with high sensitivity without any cross‐reactivity for each type through uniplex E6/E7 PCR. We confirmed a higher prevalence of multiple infections than reported previously 8 . However, PCR‐based HPV tests are always affected by false‐positives.…”

Section: Discussionsupporting

confidence: 88%

Koilocytic changes are not elicited by human papillomavirus genotypes with higher oncogenic potential

Okodo

Okayama

Teruya

et al. 2020

Journal of Medical Virology

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Koilocytes are considered a common cytopathological effect in patients with human papillomavirus (HPV) infection. Thus, we aimed to elucidate whether koilocytes are common to all HPV infections. Liquid‐based cytology samples from 651 patients with abnormal Papanicolaou (Pap) test results were used to analyze the presence of koilocytes and HPV genotype. HPV genotype was determined in complete liquid cytology samples and microdissected cell samples from Pap smear slides using the uniplex E6/E7 polymerase chain reaction method, which can detect 39 mucosal HPV genotypes. Koilocytes were found in 29.3% (191) of all patients. Logistical regression analysis of diverse HPV genotypes revealed that infections with low‐risk HPV types (HPV‐6b, HPV‐40, HPV‐42, HPV‐61, HPV‐74, HPV‐89, and HPV‐90), probably high‐risk HPV types (HPV‐53 and HPV‐66), and high‐risk types (HPV‐39 and HPV‐56) were significantly associated with the presence of koilocytes. However, HPV‐16, HPV‐18, and HPV‐52, which have higher oncogenic potential, were not found to be associated with koilocytes. These results were confirmed by HPV genotyping using microdissected koilocytes in 27 patients.Most common high‐risk types belonging to α‐9 and α‐7 genotypes appear to rarely induce koilocytic changes. Therefore, koilocytes may provide additional useful information for predicting the risk of progression to high‐grade lesions.

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Section: Discussionsupporting

confidence: 88%

Koilocytic changes are not elicited by human papillomavirus genotypes with higher oncogenic potential

Okodo

Okayama

Teruya

et al. 2020

Journal of Medical Virology

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“…The cells were assayed with Uniplex E6/E7 polymerase chain reaction (PCR), a new HPV‐typing assay method previously established byOkodo et al . The PCR mixture included 1× AmpliTaq Gold ® 360 buffer, 2 mmol/L MgCl 2 , 0.025 U/μL AmpliTaq Gold 360 DNA Polymerase (Applied Biosystems), 2.5 μL DNA, and 0.5 pmol/L primers in a total volume of 25 μL.…”

Section: Methodsmentioning

confidence: 99%

“…The cells were assayed with Uniplex E6/E7 polymerase chain reaction (PCR), a new HPV-typing assay method previously established byOkodo et al 10 The PCR mixture included 1× AmpliTaq…”

Section: Hpv Genotypingmentioning

confidence: 99%

“…High-risk HPV was detected in 63.4% (26/41) of ASC-US cases,10.5% (2/19) of NILM cases with Candida infection, and 8.8% (6/68) of NILM cases without Candida infection.Table 2shows the relationship between nuclear size and nuclear contour thickening in squamous cells with enlarged nuclei in high-risk HPV positive cases. The area ratio was 2.51 in high-risk HPV positive cases but 2.32 in negative cases.…”

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confidence: 99%

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A study of the relationship between nuclear contour thickening, nuclear enlargement and human papillomavirus infection in squamous cells

et al. 2019

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Objective To investigate the relationship between the morphological features of nuclear enlarged cells and human papillomavirus (HPV) infection in negative for intraepithelial lesion or malignancy (NILM) and atypical squamous cells of undetermined significance (ASC‐US). Methods In total, 128 Papanicolaou specimens comprising 41 ASC‐US cases and 87 NILM cases were examined. Cell morphological analysis was performed using both area ratio (nuclear area in cells with nuclear enlargement/nuclear area in normal intermediate cells) and nuclear contour thickening. High‐risk HPV was detected using the Uniplex E6/E7 polymerase chain reaction assay and logistic regression analyses of factors related to high‐risk HPV infection were performed. Results Nuclear contour thickening was present in 57.7% (64/111 cells) of high‐risk HPV positive cases and 21.8% (69/317 cells) of high‐risk HPV negative cases. There was a statistically significant association (P = 0.01) between high‐risk HPV infection and nuclear contour thickening. Nuclear contour thickening was approximately one‐third higher in NILM cases than in ASC‐US cases (odds ratio, 0.371; 95% confidence interval, 0.208–0.662) and three times higher in high‐risk HPV‐positive cases than in high‐risk HPV‐negative cases (odds ratio, 2.831; 95% confidence interval, 1.591–5.039). Conclusion Our results suggest that nuclear contour thickening in nuclear enlarged cells in NILM and ASC‐US cases may be a cellular finding associated with HPV infection.

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“…The Uniplex E6/E7 polymerase chain reaction (PCR) assay, established by Okodo et al, (2018) was used for HPV typing. DNA from the residual liquid of cervical cytology specimens (100 μL) was extracted using hot sodium hydroxide and pH adjustment with a Tris solution (HotSHOT) method (Truett et al, 2000).…”

Section: Hpv Genotypingmentioning

confidence: 99%

A Comparison of Cytomorphological Features of ASC-H Cells Based on Histopathological Results Obtained from a Colposcopic Target Biopsy Immediately after Pap smear Sampling

Hata

Okayama

Iijima

et al. 2019

Asian Pac J Cancer Prev

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Background: To compare the cytomorphological features of atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H) observed in a liquid-based Pap smear with the histopathological features observed in a concurrent colposcopic biopsy specimen obtained immediately after obtaining the Pap smear. Methods: Cytomorphological features such as cytoplasmic differentiation, nuclear/cytoplasm (N/C) ratio, chromatin pattern, thickening of nuclear contour, and the appearance of the nucleolus of 247 ASC-H obtained from 25 liquid-based Pap smear ASC-H cases were compared with those of the cells obtained from biopsied samples. Human papillomavirus (HPV) infection was tested for 39 HPV genotypes using Uniplex E6/E7 polymerase chain reaction method. Results: Of the 25 ASC-H cases, 22 (88%) showed cervical intraepithelial neoplasia grade 1 or greater (CIN1+) and 3 (12%) were benign. HPV infection was detected in 100% CIN1+ cases and 66.7% benign cases. Significant differences such as marked hyperchromasia, thickened nuclear contour, and prominent nucleoli were observed between ASC-H cases with CIN1+ and the benign cases. Conclusion: The presence of small dysplastic cells displaying marked hyperchromasia, thickening of nuclear contour, and prominent nucleoli on Pap smear strongly suggest the presence of CIN in ASC-H cases.

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Uniplex E6/E7 PCR method detecting E6 or E7 genes in 39 human papillomavirus types

Cited by 31 publications

References 23 publications

Koilocytic changes are not elicited by human papillomavirus genotypes with higher oncogenic potential

Koilocytic changes are not elicited by human papillomavirus genotypes with higher oncogenic potential

A study of the relationship between nuclear contour thickening, nuclear enlargement and human papillomavirus infection in squamous cells

A Comparison of Cytomorphological Features of ASC-H Cells Based on Histopathological Results Obtained from a Colposcopic Target Biopsy Immediately after Pap smear Sampling

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