To elucidate oncogenic human papilloma virus (HPV) types in Japan, HPV genotyping was performed in 1526 cervical intraepithelial neoplasia (CIN) and 371 invasive cervical cancer (ICC) patients with the novel Genosearch-31+5 HPV test. The HPV-positive rates were 89.3% and 90.8% in CIN and ICC. Regarding single-type infections, 13 internationally recognized high-risk (13HR) types excluding HPV 35, and probably HR HPV 53, 67, 69, and 70 were identified in ICC, suggesting that all these types may be oncogenic. HPV16 and 18 were identified in both SCC and adenocarcinoma (ADC). HPV HPV52, 31 and 58 (alpha-9) were predominantly detected in SCC, whereas HPV 18, 45, 39 and 59 (alpha-7) were in ADC. The prevalence of HPV 18 in SCC significantly decreased with increasing age of patients, whereas the opposite trend was observed in the other HR types. HPV18 is likely to induce SCC rapidly. All ICC cases aged 20–29 were positive for HPV 16 or 18, suggesting that present HPV 16, 18 vaccines may be quite effective to prevent ICC in young women.
We have developed a new human papillomavirus (HPV) assay using the uniplex E6/E7 polymerase chain reaction (PCR) method, which is able to detect E6 and E7 genes in 39 HPV types. We validated the assay for sensitivity and specificity using cloned HPV DNA and clinical samples. A comparative study using Genosearch-31 (GS-31) to determine HPV genotypes in clinical samples was also performed. E6 or E7 genes, measured by uniplex E6/E7 PCR, were detectable in 15 low-risk (HPV-6, -11, -40, -42, -44, -54, -55, -61, -62, -71, -74, -81, -84, -89, -90), 11 intermediate-risk (HPV-26, -30, -34, -53, -66, -67, -69, -70, -73, -82, -85), and 13 high-risk (HR) HPV types. The detection limit of this assay was 100 copies in all 39 HPV types and no cross-reactivity was observed with any type. This assay detected HPV in all 226 cervical cell samples, including 222 high-grade squamous intraepithelial lesions (HSIL) and 4 squamous cell carcinoma (SCC) cases, whereas GS-31 identified HPV in 99.6% (225/226) of the same samples. All SCC and 41.0% (90/222) of HSIL cases were infected with a single HPV type, while the remaining 59% of HSIL cases involved multiple HPV types. It was noted that high-risk and probably high-risk HPV types (HPV-66, -70 and -82) were identified, but no low-risk types were identified as a single-type infection in these HSIL and SCC cases. The uniplex E6/E7 PCR assay has high sensitivity, and can be useful tool in epidemiological studies or clinical follow-ups after surgery.
Background Human papillomavirus (HPV) is a well‐established mucosotropic carcinogen, but its impact on urothelial neoplasm is unclear. We aimed to clarify the clinical and pathological features of HPV‐related urothelial carcinoma (UC). Methods Tissue samples of 228 cases of UC were obtained from the bladder, upper and lower urinary tract, and metastatic sites to construct a tissue microarray. The samples were analyzed for the presence of HPV by a highly sensitive and specific mRNA in situ hybridization (RISH) technique (RNAscope) with a probe that can detect 18 varieties of high‐risk HPV. We also conducted immunohistochemistry (IHC) for a major HPV capsid antibody and DNA‐PCR. Results The HPV detection rates varied among the methods; probably due to low HPV copy numbers in UC tissues and the insufficient specificity and sensitivity of the IHC and PCR assays. The RISH method had the highest accuracy and identified HPV infection in 12 (5.2%) of the cases. The histopathological analysis of the HPV‐positive UC showed six cases of usual type UC, five cases of UC with squamous differentiation (UC_SqD), and one case of micropapillary UC. The HPV detection rate was six‐fold higher in the cases of UC_SqD than in the other variants of UC (odds ratio [OR] =8.9, p = 0.002). In addition, HPV infection showed a significant association with tumor grade (OR =9.8, p = 0.03) and stage (OR =4.7, p = 0.03) of UC. Moreover, the metastatic rate was higher in HPV‐positive than in negative UC (OR =3.4). Conclusion These data indicate that although the incidence of HPV infection in UC is low, it is significantly associated with squamous differentiation and poor prognosis. Furthermore, our observations show that RNAscope is an ideal method for HPV detection in UC compared with the other standard approaches such as IHC and PCR assays.
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.
Human papillomavirus (HPV) testing with cytology triage for cervical cancer screening has proven to be useful. It is considered that a significant percentage of HPV‐positive women followed by reflex cytology have had multiple‐type HPV infections rather than single‐type infections. However, the effects of multiple‐type infections on changes in the cytomorphology of exfoliated cervical cells have not been investigated. The aim of this study was to validate simple manual microdissection (MMD) maneuver and investigate the HPV infection status of single cells isolated from Papanicolaou (Pap) smears prepared from women with multiple‐type infections. Using cytology samples from 90 patients with abnormal Pap smear results, we evaluated the efficiency of the MMD procedure and determined the HPV infection status of single squamous intraepithelial lesion (SIL) cells microdissected from patients with multiple‐type infection. When validating the MMD procedure, the HPV‐positive rate was 81.5% using 119 MMD samples from the Pap smear in 61 cases with single‐type infection. This MMD procedure was able to efficiently collect single cells. Of 119 MMD samples from 29 cases with multiple‐type infection, the HPV‐positive rate was 42.9%, and most (96.1%) MMD samples exhibited only one genotype. Our MMD maneuver successfully identified HPV genotypes using single cells isolated from cytology specimens. A majority of single SIL cells prepared from multiple‐type infection cases turned out to contain only one genotype. In the future, the MMD method could be applied while studying the relationship between the morphological changes exhibited by SIL cells on Pap smear and the infected HPV genotype.
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