Human Papillomavirus DNA Chip Research Articles

Human papillomavirus genotyping for predicting disease progression in women with biopsy-negative or cervical intraepithelial neoplasia grade 1 of low-grade intraepithelial lesion cytology

ObjectiveOur study used human papillomavirus (HPV) genotyping to assess the disease occurrence probability in women with a low-grade squamous intraepithelial lesion (LSIL) without histologically confirmed cervical intraepithelial neoplasia grade 2...

Usefulness of the human papillomavirus DNA chip test as a complementary method for cervical cytology.

As a convenient and economical method of screening cervical cancer and precancerous pathologies, the Papanicolaou smear (Pap smear) has been most widely used. Nevertheless, it requires cytological changes for making diagnoses and reportedly has a high false-negative rate. In this study, the usefulness of the human papillomavirus (HPV) DNA chip test as a complementary method that can compensate for the defect of the Pap smear was investigated. Of the 6516 patients who simultaneously underwent a Pap smear and an HPV DNA chip test at Chonnam National University Hospital between January 2015 and December 2016, 1897, an initial PAP smear-negative patients who had undergone an additional Pap smear during their 2-year follow-up period were selected for this study. Of the subject patients, 281 underwent a cervical biopsy. The Pap smear follow-up of an initial Pap smear-negative subjects showed 53 (75.7%) HPV high-risk positive cases in the cytology low-grade lesion group (70 cases) and 46 (97.8%) HPV high-risk positive cases in the cytology high-grade lesion group (47 cases). The 281 biopsy cases included 67 biopsy low-grade lesion cases and 74 biopsy high-grade lesion cases, of which there were 45 (67.2%) and 67 (90.5%) HPV high-risk positive cases, respectively. The follow-up cytology on the high-risk HPV-positive subjects showed that the ratio of their high-grade lesions was 260.8 times greater than that of the high-risk HPV-negative subjects (OR = 260.8 and 95% CI: 36.1 and 1886.1); and their biopsy showed that the ratio of their high-grade lesions was 102.7 times greater than that of the HPV-negative subjects (OR = 102.7 and 95% CI: 14.0 and 753.3). The complementary use of the HPV DNA chip test may be useful in increasing the accuracy of screening examinations for the early diagnosis of uterine cervix cancer when combined with the Pap smear.

Predictive value of HPV genotyping for recurrence of CIN2−3 in women treated by LEEP with negative resection margins

The aim of our study was to determine whether the HPV genotype identified by the HPV DNA chip (HDC) test could predict the recurrence of high-grade cervical intraepithelial neoplasia (CIN2–3) in women who had undergone a loop electrosurgical excision procedure (LEEP) and had negative margins. We analyzed the data of 1021 women with CIN2–3 treated by LEEP where histology confirmed negative resection margins. The women were followed up with HDC and endocervical cytology tests at 3, 6, 9, 12, 18 and 24 months during the first 2 years and annually thereafter. Among the 1021 patients, the pre-LEEP HDC test was positive for 992 (97.2%). A total of 90 (8.8%) patients experienced recurrence of CIN2–3 and the post-LEEP follow-up HDC tests were positive, thus demonstrating a persistent high-risk HPV infection of the same genotype, which showed a sensitivity and negative predictive value of 100%in predicting recurrence. We also examined the correlation between pre-LEEP high-risk HPV genotypes and recurrence. The most common subtypes were HPV16, HPV18 and HPV31. Persistence of HPV18 had the highest risk of recurrence of CIN2–3 (p < 0.05). Our study suggests that type-specific persistent high-risk HPV infection, particularly the HPV18 subtype, is a significant predictor of recurrence in patients with negative resection margins in the LEEP specimens. Thus, these patients require careful monitoring, and gynecologic oncologists should conduct short-term follow-up tests.

Comparison of the performance of HPV DNA chip test and HPV PCR test in cervical cancer screening in rural China.

This study aimed to evaluate the performance of two different principles of HPV testing in primary cervical cancer screening and ASC-US triage in rural areas. 3,328 and 3,913 women were enrolled in Shanxi, China in 2017 and 2018, respectively, and screened using liquid-based cytology and different HPV tests with a 4-year follow-up. Different screening methods commonly used in clinical practice were evaluated. In the HPV PCR test cohort, the prevalence of HPV infection was 14.90%. A total of 38 cases of CIN2+ were identified at baseline, 2 of which were in the HPV-negative cohort and the rest in the HPV-positive cohort (2 = 186.85, p < 0.001). Fifty-three cases of CIN2+ were accumulated over 4 years. The HPV infection rate in the HPV DNA chip test cohort was 21.10%. A total of 26 CIN2+ cases were identified at baseline, all in the HPV-positive population (2 = 92.96, p < 0.001). 54 CIN2+ cases were cumulative over 4 years. At 4-year follow-up, HPV-negative results were significantly more protective against cervical intraepithelial neoplasia grade 2 or worse (CIN2+) than normal cytologic results at baseline. HPV screening was more sensitive and specific than cytologic screening (using ASC-US as the threshold) and performed better on the HPV DNA microarray test. In addition, compared with HPV 16/18 testing, sensitivity increases and specificity decreases when using HPV testing for cytologic ASC-US triage, regardless of which HPV test is used. In the rural areas where we implemented the study, HPV tests performed well for screening than LBC and HPV DNA chip testing performed better than HPV PCR testing in the screening cohort. Optimal screening was achieved technically when used in combination with LBC for ASC-US population triage, without thinking the feasibility for resource availability.

Human papillomavirus genotype-specific risk in cervical carcinogenesis.

ObjectiveTo evaluate the risk of genotype-specific human papillomavirus (HPV) infections for the spectrum of cervical carcinogenesis and the distribution of HPV types according to age and different cervical lesionsMethodsThis study included HPV-positive women who underwent cervical biopsy at the Cheil General Hospital & Women's Healthcare Center between July 1, 2011 and December 31, 2017. HPV genotyping was conducted using a Cheil HPV DNA chip kit.ResultsThe study sample consisted of 400 normal, 399 cervical intraepithelial neoplasia (CIN) 1, 400 CIN 2, 400 CIN 3, and 389 cervical cancer cases. HPV 16 was the most common type found with a prevalence of 9.5% in normal, 6.8% in CIN 1, 15.0% in CIN 2, 44.5% in CIN 3, and 64.3% in cervical cancer. The most common HPV types were 16, 52, 58, 53, 51, 56, 68, and 18 in all study samples. HPV 16, 31, 33, and 58 were more common in CIN 2/3 and cancer, and HPV 39, 51, 53, 56, 66, and 68 were more common in CIN 1 and normal cases (p<0.001). In CIN 3 and cervical cancer, HPV 16 was the most common type in all age groups. HPV 52 was the most common type in CIN 2 (all age groups) and in CIN 1/normal (age ≤30 years) cases. Among the high-risk HPV types, 16, 31, 33, 52, and 58 showed significant risk for high-grade disease.ConclusionsHPV 16, 31, 33, 52, and 58 showed the significant risk of high-grade disease for cervical carcinogenesis.

Poor prognostic factors in human papillomavirus-positive head and neck cancer: who might not be candidates for de-escalation treatment?

Background/AimsSince patients with human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) have favorable outcomes after treatment, treatment de-escalation for these patients is being actively investigated. However, not all HPV-positive HNSCCs are curable, and some patients have a poor prognosis. The purpose of this study was to identify poor prognostic factors in patients with HPV-positive HNSCC.MethodsPatients who received a diagnosis of HNSCC and tested positive for HPV from 2000 to 2015 at a single hospital site (n = 152) were included in this retrospective analysis. HPV typing was conducted using the HPV DNA chip assay or liquid bead microarray system. Expression of p16 in the tumors was assessed by immunohistochemistry. To determine candidate factors associated with overall survival (OS), univariate and multivariable Cox regression analyses were performed.ResultsA total of 152 patients with HPV-positive HNSCC were included in this study; 82.2% were male, 43.4% were current or former smokers, and 84.2% had oropharyngeal cancer. By univariate analysis, old age, performance status ≥ 1, non-oropharyngeal location, advanced T classification (T3–4), and HPV genotype 18 were significantly associated with poor OS. By multivariable analysis, performance status ≥ 1 and non-oropharyngeal location were independently associated with shorter OS (hazard ratio [HR], 4.36, p = 0.015; HR, 11.83, p = 0.002, respectively). Furthermore, HPV genotype 18 positivity was also an independent poor prognostic factor of OS (HR, 10.87, p < 0.001).ConclusionsNon-oropharyngeal cancer, poor performance status, and HPV genotype 18 were independent poor prognostic factors in patients with HPV-positive HNSCC. Patients with these risk factors might not be candidates for de-escalation treatment.

A Review and Update on Papillary Immature Metaplasia of the Uterine Cervix: A Distinct Subset of Low-Grade Squamous Intraepithelial Lesion, Proposing a Possible Cell of Origin.

- Papillary immature metaplasia (PIM) is a known papillary cervical lesion associated with low-risk human papillomavirus (LR-HPV). - To evaluate additional clinicopathologic features and the HPV genotypes of PIM and discuss the presumptive cell of origin. - A total of 26 PIM cases were evaluated by p16INK4a, cytokeratin (CK) 7, and CK17 immunohistochemical stainings. Human papillomavirus genotyping was performed, by using HPV DNA Chip, HPV polymerase chain reaction (PCR), and real-time PCR. - Histologically, PIM forms either a papillary mass (n = 21 of 26, 81%) or a slightly elevated/flat plaque (n = 5, 19%). All cases contain variable amounts of mucinous epithelia within the lesions. Koilocytosis was identified in 15 of the 26 cases (58%). Sixteen cases (61%) were associated with LR-HPV (types 6, 11, or 42), but 3 cases (12%) with high-risk (HR) HPV (16, 16/18, and 33), 2 cases (8%) with mixed LR- and HR-HPV (6/16 and 11/58), while 2 cases (8%) were negative, but p16INK4a immunostaining showed nonblock positivity in all cases. Eight (31%) had high-grade squamous intraepithelial lesion (HSIL) in the adjacent mucosa, 4 (50%) of which showed direct continuity. Identical HPV subtypes were confirmed in separately microdissected cases from PIM and adjacent HSIL. Most lesions (n = 24, 92%) expressed CK17 (reserve cell marker) in a bottom-heavy pattern and CK7 (squamocolumnar junction [SCJ] marker) in a top-heavy pattern, while most cases of low-grade squamous intraepithelial lesion (LSIL) were negative for both markers. - Our results suggest that PIM is a distinct subset of LSIL showing a productive HPV infection, but PIM involves the transformation zone and is proximal to SCJ, while LSIL is mostly from ectocervix or distal to the SCJ.

Sequencing analysis of HPV-other type on an HPV DNA chip.

ObjectivesTo identify the specific human papillomavirus (HPV) genotypes from HPV-other type on an HPV DNA chip test by sequencing.MethodsAmong 13,600 women undergoing a routine gynecology examination including Pap smear and/or HPV test by DNA chip test in the healthcare system at Gangnam Center from July 2012 to February 2013, we prospectively collected and performed sequencing for a total of 351 consecutive cervicovaginal samples consisting of 180 samples that tested positive for HPV-other type and 171 samples that tested positive for either high-risk HPV or low-risk HPV.ResultsOf a total of 351 samples, individual HPV genotypes were successfully sequenced in 215 cases: 119 HPV-other type, 82 HPV-high-risk, and 14 HPV-low-risk. Based on the sequencing for 119 HPV-other type samples, 91.6% were detected as HPV types that were not included on the DNA chip; however, 7.6% (9/119) were proven to be high-risk HPV types: HPV 18 (n=4), HPV 33 (n=3), HPV 35 (n=1), and HPV 59 (n=1). For correlation analysis of all high-risk and HPV 16/18, the correlation rate was 76.2% and 86.6% with kappa-value of 0.38 and 0.69, respectively.ConclusionHPV-other type on DNA chip test may still have possibility of high-risk HPV, i.e., HPV 18 and thus the significance of HPV-other type in detecting cervical disease remains to be investigated.

Is human papillomavirus genotype important in predicting disease progression in women with biopsy-proven negative or CIN1 of atypical squamous cell of undetermined significance (ASC-US) cytology?

ObjectivesOur aim was to estimate the risk of disease incidence in women with atypical squamous cell of undetermined significance (ASC-US) without histology-proven cervical intraepithelial neoplasia grade 2 or worse (CIN2+) by human papillomavirus (HPV) genotype. MethodsBetween January 2002 and September 2010, incidence of CIN2+ in 2880 women including 2172 with ASC-US and histology-proven negative and 708 with ASC-US with histology-proven CIN1 was investigated. Baseline HR-HPV status was determined by the hybrid capture II assay (HC2) and HR-HPV genotype by the HPV DNA chip test (HDC). Cumulative incidence and hazard ratios were estimated to explore differences between index data and associations with CIN2+. ResultsOf the 2880 women, the HC2 was positive in 1509 women (52.4%) and the HDC was positive in 1563 women (54.3%). The overall agreement between the HDC and HC2 was 97.4%. One hundred ninety (6.6%) patients developed CIN2+. The 5-year cumulative incidence rate of CIN2+ in HPV-16, HPV-31, HPV-52, and HPV-58 were 16.7%, 15.1%, 12.6%, and 12.9%, respectively. On multivariate analysis, being positive in HPV-16 (hazards ratio [HR]=2.431; 95% CI, 1.789–3.332; P<0.01), HPV-31 (HR=2.335; 95% CI, 1.373–3.971; P<0.01), HPV-52 (HR=1.592; 95% CI, 1.031–2.458; P=0.03), and HPV-58 (HR=1.650; 95% CI, 1.132–2.407; P<0.01) were significantly associated with developing CIN2+ compared to being negative for that type. ConclusionsAmong women with ASC-US, HPV-16, HPV-31, HPV-52, or HPV-58 positive women may need intensified follow-up as they have the highest risk of becoming CIN2+.

P16-negative warty carcinoma of the uterine cervix with superficial invasion to the endometrium: A case report focusing on diagnostic pitfalls

Warty carcinoma of the uterine cervix is a very rare and specific variant of invasive squamous cell carcinoma, usually described as a hybrid of the features of both condyloma acuminatum and invasive squamous cell carcinoma. The diagnostic pitfalls of this lesion are: 1) the bland appearance of cytomorphology, 2) distinct koilocytotic atypia, and 3) p16 negativity. A case of warty carcinoma with a fungating mass on the uterine cervix of a 75-year-old woman is presented. The original diagnosis by punch biopsy was atypical squamous cells, undetermined for malignancy, adjunct with p16 negativity. The subsequent radical hysterectomy showed a protruding mass (5.7 cm in horizontal dimension) of the uterine cervix. Light microscopy revealed a protruding mass composed of exophytic papillae and inverted nests along the endocervical glands. There were multiple micro-invasive foci (&lt; 1 mm) of tongue like projections at the nest base and slight involvement of the upper vagina and endometrial surface. The p16 immunohistochemical staining yielded negative results. The results of the HPV DNA chip test from paraffin-embedded tissue were 6 (low risk, +++) and 42 (low risk, +). The Ki-67 proliferation index was approximately 20%. The invasive foci and patient’s age were the determining differential factors for malignancy rather than benign condylomatous lesion.

Clinical progress of human papillomavirus genotypes and their persistent infection in subjects with atypical squamous cells of undetermined significance cytology: Statistical and latent Dirichlet allocation analysis.

The present study aimed to investigate differences in prognosis based on human papillomavirus (HPV) infection, persistent infection and genotype variations for patients exhibiting atypical squamous cells of undetermined significance (ASCUS) in their initial Papanicolaou (PAP) test results. A latent Dirichlet allocation (LDA)-based tool was developed that may offer a facilitated means of communication to be employed during patient-doctor consultations. The present study assessed 491 patients (139 HPV-positive and 352 HPV-negative cases) with a PAP test result of ASCUS with a follow-up period ≥2 years. Patients underwent PAP and HPV DNA chip tests between January 2006 and January 2009. The HPV-positive subjects were followed up with at least 2 instances of PAP and HPV DNA chip tests. The most common genotypes observed were HPV-16 (25.9%, 36/139), HPV-52 (14.4%, 20/139), HPV-58 (13.7%, 19/139), HPV-56 (11.5%, 16/139), HPV-51 (9.4%, 13/139) and HPV-18 (8.6%, 12/139). A total of 33.3% (12/36) patients positive for HPV-16 had cervical intraepithelial neoplasia (CIN)2 or a worse result, which was significantly higher than the prevalence of CIN2 of 1.8% (8/455) in patients negative for HPV-16 (P<0.001), while no significant association was identified for other genotypes in terms of genotype and clinical progress. There was a significant association between clearance and good prognosis (P<0.001). Persistent infection was higher in patients aged ≥51 years (38.7%) than in those aged ≤50 years (20.4%; P=0.036). Progression from persistent infection to CIN2 or worse (19/34, 55.9%) was higher than clearance (0/105, 0.0%; P<0.001). In the LDA analysis, using symmetric Dirichlet priors α=0.1 and β=0.01, and clusters (k)=5 or 10 provided the most meaningful groupings. Statistical and LDA analyses produced consistent results regarding the association between persistent infection of HPV-16, old age and long infection period with a clinical progression of CIN2 or worse. Therefore, LDA results may be presented as explanatory evidence during time-constrained patient-doctor consultations in order to deliver information regarding the patient's status.

Comparison of FFPE histological versus LBP cytological samples for HPV detection and typing in cervical cancer

Human papillomavirus (HPV) infection is closely associated with cervical cancer. This study analyzed HPV genotype prevalence in 75 cases of formalin-fixed paraffin embedded (FFPE) tissue samples from patients diagnosed with cervical cancer. Genotype prevalence was assessed using Reverse Blot Assay (REBA) and quantitative polymerase chain reaction (qPCR), which target the HPV L1 and HPV E6/E7 genes, respectively. HPV DNA chip tests were also performed using liquid based preparation (LBP) cytological samples from the same patients who provided the FFPE histological samples. We observed a slight difference in HPV genotype distribution as assessed by DNA chip versus REBA. One possible explanation for this difference is that normal regions could be mixed with lesion regions when cytological samples are extracted from each patient with cancer. For the detection of moderate dysplasia, the main target of diagnosis, this difference is anticipated to be greater. We also made several unexpected observations. For example, HPV multi-infection was not detected. Moreover, the rate of HPV positivity varied radically depending on the cancer origin, e.g. squamous cell carcinoma versus adenocarcinoma.Our results imply that it is important to determine whether cytological specimens are suitable for HPV genotyping analysis and cervical cancer diagnosis. Future research on the mechanisms underlying cervical cancer pathogenesis is also necessary.

Clinical Significance of an HPV DNA Chip Test with Emphasis on HPV-16 and/or HPV-18 Detection in Korean Gynecological Patients.

Background:Human papillomavirus (HPV) is a major risk factor for cervical cancer.Methods:We evaluated the clinical significance of the HPV DNA chip genotyping assay (MyHPV chip, Mygene Co.) compared with the Hybrid Capture 2 (HC2) chemiluminescent nucleic acid hybridization kit (Digene Corp.) in 867 patients.Results:The concordance rate between the MyHPV chip and HC2 was 79.4% (kappa coefficient, κ = 0.55). The sensitivity and specificity of both HPV tests were very similar (approximately 85% and 50%, respectively). The addition of HPV result (either MyHPV chip or HC2) to cytology improved the sensitivity (95%, each) but reduced the specificity (approximately 30%, each) compared with the HPV test or cytology alone. Based on the MyHPV chip results, the odds ratio (OR) for ≥ high-grade squamous intraepithelial lesions (HSILs) was 9.9 in the HPV-16/18 (+) group and 3.7 in the non-16/18 high-risk (HR)-HPV (+) group. Based on the HC2 results, the OR for ≥ HSILs was 5.9 in the HR-HPV (+) group. When considering only patients with cytological diagnoses of “negative for intraepithelial lesion or malignancy” and “atypical squamous cell or atypical glandular cell,” based on the MyHPV chip results, the ORs for ≥ HSILs were 6.8 and 11.7, respectively, in the HPV-16/18 (+) group.Conclusions:The sensitivity and specificity of the MyHPV chip test are similar to the HC2. Detecting HPV-16/18 with an HPV DNA chip test, which is commonly used in many Asian countries, is useful in assessing the risk of high-grade cervical lesions.

Smoking, not human papilloma virus infection, is a risk factor for recurrence of sinonasal inverted papilloma.

The recurrence rate of sinonasal inverted papillomas (SNIP) is 15-20%. However, few studies have investigated patient-dependent factors related to recurrence of SNIPs. To analyze risk factors, including human papilloma virus (HPV) infection and smoking, as well as other factors, for recurrence of SNIPs. Fifty-four patients who were diagnosed with SNIP and underwent surgery were enrolled: 39 men and 15 women, with the mean age of 54.0 years. Their mean follow-up was 40.6 months. Demographics and information about the history of smoking, previous surgery, tumor extent, follow-up, and recurrence were reviewed retrospectively. Those patients whose tumors were associated with malignant transformation were excluded in this study. HPV detection and genotyping in the tumor specimens were performed with the HPV DNA chip, a polymerase chain reaction-based DNA microarray system. Seven patients (13.0%) had recurrence, with a mean time to recurrence of 39.8 months. Recurrence rates in T1, T2, T3, and T4 of the Krouse staging system were 0% (0/4), 8.3% (2/24), 17.4% (4/23), and 33.3% (1/3), respectively (p > 0.5). Eight patients (14.8%) were positive for HPV DNA. All of these patients belonged to the group without recurrence (p > 0.5). However, recurrence rates according to HPV DNA positivity were not statistically different (0% versus 15.2%). Three (42.9%) in the group with recurrence and four (8.5%) in the group without recurrence were smokers (p < 0.5). Smoking was associated with recurrence of SNIP. However, HPV infection is not a recurrence of SNIP risk factor.

Comparison of Analytical and Clinical Performance of HPV 9G DNA Chip, PANArray HPV Genotyping Chip, and Hybrid-Capture II Assay in Cervicovaginal Swabs

Background:Human papillomavirus (HPV) infection can be detected by using several molecular methods, including Hybrid-Capture II (HC2) assay and variable HPV DNA chip tests, although each method has different sensitivities and specificities.Methods:We performed HPV 9G DNA Chip (9G) and PANArray HPV Genotyping Chip (PANArray) tests on 118 cervicovaginal swabs and compared the results with HC2, cytology, histology, and direct sequencing results.ResultsThe overall and high-risk HPV (HR-HPV) positivity rates were 62.7% and 44.9% using 9G, and 61.0% and 30.5% using PANArray, respectively. The positivity rates for HR-HPV with these two chips were significantly lower than 55.1% when HC2 was used. The sensitivity of overall HPV positivity in detecting histologically confirmed low-grade cervical squamous intraepithelial lesions or higher was 88.7% for all three tests. The specificity was 58.5% for 9G and 61.5% for PANArray, which was significantly lower than the 72.3% for HC2. With the HR-HPV+ genotype threshold, the sensitivity decreased to 75.5% for 9G and 52.8% for PANArray, which was significantly lower than the 88.7% for HC2. Comparison of the two chips showed concordant results in 55.1% of the samples, compatible results in 16.9%, and discordant results in 28.0%, exhibiting poor agreement in detecting certain HPV genotypes. Compared with direct sequencing, 9G yielded no discordant results, whereas PANArray yielded 31 discordant results (26.7%).ConclusionsCompared with HC2, the HPV genotyping tests showed lower sensitivity in histologic correlation. When the two chips were compared, the 9G was more sensitive and accurate for detecting HR-HPV than the PANArray.

Human papillomavirus genotyping as a reliable prognostic marker of recurrence after loop electrosurgical excision procedure for high-grade cervical intraepithelial neoplasia (CIN2-3) especially in postmenopausal women.

This study was conducted to determine, using the HPV DNA Chip (HDC) test, whether the human papillomavirus (HPV) genotype is predictive of recurrent high-grade cervical intraepithelial neoplasia (CIN; CIN2-3) after a loop electrosurgical excision procedure (LEEP) in postmenopausal women. Between January 2007 and February 2013, 206 postmenopausal women with CIN2-3 were treated with LEEP, followed by cytology, Hybrid Capture II (HC2) assay, and HDC test. Post-LEEP follow-up was performed at 3, 6, 9, 12, 18, and 24 months during the first 2 years and yearly thereafter. Among 206 women, HC2 yielded positive results in 199 women (96.6%) and HDC yielded positive results in 201 women (97.6%) before LEEP. The overall agreement between HDC and HC2 was 99.0%. The area under the receiver operating characteristic curve for high-risk HPV (HR-HPV) viral load measured by HC2 predicting recurrent CIN2-3 was 0.567 (P = 0.335). Twenty-six women (12.6%) developed recurrence, and those who developed recurrence tested positive for the same HR-HPV genotype before and after LEEP. The same HR-HPV genotype by HDC during follow-up had a sensitivity and negative predictive value of 100% in detecting recurrent disease. HPV-18 was significantly associated with recurrent CIN2-3 (P < 0.05). Among postmenopausal women, persistent infection with the same HR-HPV genotype, especially HPV-18, should be considered a risk factor for developing recurrent CIN2-3. After LEEP, such women warrant special attention with intense follow-up.

Combined SYBR Green real-time polymerase chain reaction and microarray method for the simultaneous determination of human papillomavirus loads and genotypes

ObjectiveThe aim of this study was to describe the principle of the Cheil HPV DNA Chip assay and evaluate its accuracy. In order to quantify the human papillomavirus (HPV) load and identify HPV genotypes simultaneously, this assay combined the two methods: SYBR Green quantitative real-time polymerase chain reaction (PCR) and DNA microarray.MethodsWe designed novel consensus primer sets that target the conserved region of the HPV L1 gene for quantifying and detecting a broad range of HPV types by quantitative real-time PCR. Subsequently, using the PCR products, DNA microarray was performed with 36 HPV type-specific probes. To validate this method, direct sequencing and correlation analysis among HPV genotype, viral load, and cytological abnormality was performed by Cohen’s kappa values, two-sided McNemar chi-square test, Kruskal-Wallis test, and odds ratios.ResultsThe kappa value of the Cheil HPV DNA Chip was 0.963 (95% confidence interval, 0.919 to 0.98), which was significantly higher than the value of 0.527 (95% confidence interval, 0.447 to 0.59) obtained using a conventional HPV DNA Chip. HPV16 (χ2=62.28, P<0.01), HPV33 (χ2=7.18, P<0.01), and HPV58 (χ2=9.52, P<0.01), which are classified as high-risk HPVs, were detected at significant levels in samples with high-grade lesions. And viral loads tended to be higher in groups with high odds ratios.ConclusionThe Cheil HPV DNA Chip is an effective diagnostic assay for simultaneously detecting HPV genotypes and loads in cervical samples.

A Case of Multiple Epidermolytic Acanthoma of the Scrotum: Is the Human Papillomavirus a Culprit?

Dear Editor: Epidermolytic acanthoma is a rare benign tumor that was first described in 1970 by Shaprio and Baraf. It usually presents in adulthood as an asymptomatic tumor <1 cm in diameter with a verrucous surface1. Herein, we report a very uncommon case of multiple epidermolytic acanthoma localized on the scrotum. A 60-year-old man visited our clinic with asymptomatic scrotal dermatosis that was discovered several months ago. He had no significant medical history and denied any triggering factors such as trauma, immunosuppressive therapy, or radiation exposure. Physical examination revealed multiple well-demarcated gray to whitish round papules on the scrotum (Fig. 1). Laboratory tests, including complete blood count, chemical battery, and screening tests for HIV and treponemal disease such as syphilis, were performed. No abnormal finding was found. Skin biopsy showed hyperkeratosis without parakeratosis and a hypertrophic granular layer with numerous keratohyalin granules. Separation and edematous changes in the spinous layer were also observed (Fig. 2). The histopathological findings were consistent with epidermolytic acanthoma. The differential diagnosis included epidermolytic verrucous nevus, bullous congenital ichthyosiform erythroderma, and Vorner's palmoplantar keratoderma with a rather distinct clinical aspect. Seborrhoreic keratoses, solar keratoses, or Bowen's disease could also show some focal changes of epidermolytic hyperkeratosis. We also tested for the presence of the human papillomavirus (HPV) genome by using a novel HPV DNA chip (GG HPVCHIP; Goodgene Inc., Seoul, Korea). The lesion was HPV 16 positive2. Fig. 1 Multiple well-demarcated <5 mm gray to whitish round papules on the scrotum. Fig. 2 Compact hyperkeratosis with acanthosis. Perinuclear vacuolization, of variable size, of the keratinocytes in the stratum spinosum and stratum granulosum. The keratinocytes exhibited non-well-defined limits that gave the epidermis a reticulate appearance. ... Epidermolytic acanthoma is a benign, asymptomatic tumor that usually presents as a solitary papule in adulthood. The multiple or disseminated form was also reported as a rare variant1. The pathogenesis of epidermolytic acanthoma is not clear. Various contributing factors have been reported, such as HPV, other viral infections, ultraviolet radiation, immunosuppression, trauma, and mutations of genes encoding for keratin 1 and 103. Although some authors have suggested the role of HPV in the pathogenesis, immunohistochemical and molecular studies had failed to detect a viral genome in the lesions4. However, the authors used a conventional method: polymerase chain reaction followed by dot-blot analysis with specific radiolabeled detection probes, which have limitations in detecting and typing HPV. The HPV DNA chip arrayed by using multiple oligonucleotide probes for 42 types of HPV and human β-globin gene, which we used in our case, was recently developed and has been found to be a highly sensitive tool for identifying HPV2. In our case, doubt exists about whether the presence of HPV 16 was only a coincidence, because HPV can be detected in otherwise normal genital skin. Although we could not clarify the possible pathogenic role of HPV in epidermolytic acanthoma, our case is important because it is the first reported case of HPV detected on skin affected by epidermolytic acanthoma. Although larger studies including several cases are required to prove the possibility, several facts imply the role of HPV in the pathogenesis of epidermolytic acanthoma: its site of predilection is the genital area; it usually has a verrucous surface; and, in a recent report, epidermolytic acanthomas were successfully treated with topical imiquimod, which is also used to treat HPV-induced lesions5.

Detection of Human Papillomavirus Type 39 in a Seborrheic Inclusion Cyst of the Buttock

A seborrheic inclusion cyst (SIC) is a lesion, which has been described as a combination of seborrheic keratosis and an epidermoid cyst (EC). To the best of our knowledge, only eight cases of SIC have been reported thus far in the English literature[1-3]. Microscopically, SICs exhibit features that are suggestive of viral infection[4]. Therefore, an association between SICs and human papillomavirus (HPV) has been suspected, but it has only been demonstrated in one previous report by Terada,[1] which identified an HPV-positive SIC through immunohistochemical staining for HPV antigens. However, no SIC study has ever used HPV genotyping. Herein, we report the first case of SIC associated with HPV type 39, which we identified using the HPV DNA chip.

Répartition des génotypes de papillomavirus humain dans les frottis ASC-US de patientes infectées ou non par le VIH-1. Résultats préliminaires d’une enquête rétrospective locale

ObjectivesAnalysis of the distribution of human papillomavirus (HPV) genotypes amongst HIV-negative and HIV-positive women diagnosed with atypical squamous cells of undetermined significance (ASC-US). Patients and methodsWe performed a retrospective study of HPV genotype distribution in 313 ASC-US Pap smears from women who consulted at the University Hospital of Nice between 2008 and 2012. HPV genotyping results, conducted on PapilloCheck® HPV DNA Chip, and information on the status of HIV patients were retrospectively collected. The odds ratio were calculated by logistic regression. ResultsIn co-infected HIV/HPV women we observed a low prevalence of HPV16, a high prevalence of oncogenic HPV low risk and a high prevalence of HPV 68 compared to non-co-infected women. By grouping HPV HR based on their prevalence in cervical cancer of the uterus and their genetic proximity we observe that HPV 18, 45, 68 (the “alpha-7”) are 7.4 times more represented (CI95 [2.48 to 22.35]) than HPV 16, 31, 33, 52, 58 (the “alpha-9”) in the population of women co-infected with HIV. Discussion and conclusionGiven that HPV “alpha-7” are responsible for 46.3% of adenocarcinomas, the high prevalence of these HPV found in ASC-US Pap smears of co-infected women should be put in relation with the highest prevalence of glandular abnormalities found in this population. HPV genotyping could become an essential tool for gynecological care for HIV positive women.