scholarly journals HPV Testing of Biobanked Liquid-Based Cytology – a Validation Study

2016 ◽  
Vol 31 (2) ◽  
pp. 218-223 ◽  
Author(s):  
Gabriella Lillsunde Larsson ◽  
Mats G. Karlsson ◽  
Gisela Helenius

Introduction The aim of the study was to investigate whether biobanked liquid-based cytology (LBC) vaginal samples could be reanalyzed for the biomarkers HPV DNA and mRNA without loss of sensitivity. Methods One hundred LBC samples with ASCUS or CIN1 were tested for HPV DNA and mRNA before and after biobanking. DNA analysis targeted the viral genes E6 and E7, 12 high-risk and 2 low-risk HPV types together with the human control gene HBB, using real-time PCR. The Aptima HPV assay was used for mRNA analysis of 14 high-risk HPV types. Results With Aptima there was 84% agreement between results before and after biobanking. The sensitivity and specificity were 0.79 (95% CI, 0.68-0.88) and 0.94 (95% CI, 0.80-0.99), respectively. With the DNA-based method, the agreement between results was 87%, the sensitivity 0.85 (95% CI, 0.75-0.92) and the specificity 0.95 (95% CI, 0.77-1.00). Both methods presented a significant difference between positive results before and after biobanking; McNemar test: p = 0.004, p = 0.003, Cohen's kappa: 0.67 (95% CI, 0.53-0.81), 0.68 (95% CI, 0.52-0.84). Cycle threshold values for the DNA method were higher for all genotypes after biobanking, except for HPV-59. Some loss of sensitivity was seen after biobanking but the concordance between HPV detection before and after biobanking was good for both evaluated methods. Conclusions Biobanking of LBC vaginal samples offers a good platform for HPV testing and could be extended to further molecular analyses. However, in order to ensure a valid test result a larger portion needs to be analyzed from the biobanked sample.

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 239
Author(s):  
Ashley L. Reily-Bell ◽  
Amanda Fisher ◽  
Bryony Harrison ◽  
Sara Bowie ◽  
Sankalita Ray ◽  
...  

Whether HPV is causative of pregnancy complications is uncertain. E6 and E7 affect functions underling preeclampsia (PET) in cultured trophoblasts, but whether E6 and E7 is produced in the placenta is uncertain. Here, we investigated whether E6/E7 was expressed in the placentae from pregnancies with PET, other pregnancy complications (fetal growth restriction (FGR) and diabetes mellitus), and uncomplicated pregnancies. Placental tissues collected from two geographical locations were subjected to RNAscope analyses of high- and low- risk E6/E7, and individual HPV types identified using an HPV array. High-risk E6/E7 expression was increased in both PET cohorts, (81% and 86% of patients positive for high-risk HPV DNA compared to 13% of control patients). Various HPV types were identified. Although HPV 18 was the most frequent in all cohorts, the majority of individuals had multiple HPV types (55% of the PET compared to 25% of the control cohort). Further evidence that E6 and E7 is present early when placental pathology underlying preeclampsia is established, is provided with the finding of high-risk E6/E7 in the first-trimester placenta anchoring trophoblast. In conclusion, E6/E7 expression and multiple HPV types were frequent in placentae from preeclampsia-complicated pregnancies.


2011 ◽  
Vol 152 (45) ◽  
pp. 1804-1807
Author(s):  
Ádám Galamb ◽  
Attila Pajor ◽  
Zoltán Langmár ◽  
Gábor Sobel

Human papilloma virus (HPV) is the most common sexually transmitted infection in the 21st century. It has been established that infections with specific HPV types are contributing factors to cervical cancer. Approximately 99.7% of cervical cancers are associated with high risk HPV types. HPV testing plays an important role in the prevention, by decreasing the prevalence and the mortality of cervical cancer. There are 16 HPV-centers operating in Hungary, in which patients undergo HPV screening, cervical exams, and treatment based on standardized guidelines. Patients and methods: The first HPV-center was founded in 2007 in Budapest, at the 2nd Department of Obstetrics and Gynecology, Semmelweis University. This study aimed to define the presence and prevalence of HPV-DNA in the cervical swab samples obtained from patients in our center. Authors conducted to assess the age-specific-prevalence, and HPV type distribution, the associated cervical abnormalities, comparing our results with international data. Results: Overall 1155 woman underwent HPV-testing and genotyping, using polymerase chain reaction. Overall, 55.5% of patients had positive test for HPV DNA types, in which 38.5% for high-risk HPV DNA. Overall prevalence was the highest among females aged 15 to 25years (62.9%). The most common HPV type found was the high risk type 16 (19.5% among the patients with positive HPV testing). Presence of high risk HPV with concurrent cervical cytological abnormality was in 32%. More than two-thirds of woman with cytological atypia (70.6%) were infected with two or more high risk HPV types. HPV 16 was detected in 32% of patients with cytological abnormalities. Conclusions: The results suggest that the prevalence of HPV in this study population exceeds the international data. The results attracts the attention the peak prevalence of the high risk types in the youngest age-group, and the higher risk of cervical abnormality in case of presence of two or more HPV types. The dominance of type 16 and 18 was predictable, but the strong attendance of type 51 and 31 among patients who had cytological atypia, was slightly surprising. Orv. Hetil., 2011, 152, 1804–1807.


Author(s):  
Junita Indarti ◽  
Darrell Fernando ◽  
Finish Fernando ◽  
Ribkhi A. Putri ◽  
Anggara Mahardika ◽  
...  

Abstract Objective: This research aim is to report the incidence of positive HPV high risk in negative cytology result. Method: We collected 83 women underwent liquid-based cytology (LBC) and HPV DNA examination at the same time. We were using DiagCor GenoFlow Human Papillomavirus Array Test (GenoFlow), a novel HPV test based on PCR and "Flow-through" hybridization that can identify 33 HPV subtypes: 18 types of High risk HPV such as 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 81 and 82 Result: We grouped the subjects based on age below or equal to 30 years old (n=6) and above 30 years old (n=77). We found a significant difference in HPV DNA result within this group (P = 0.034), with 19.3% had HPV DNA type 16 and 18 in a group of age above 30 years old. Our study showed that 27 women (32.5%) underwent screening for cervical cancer having negative LBC result but showed positive HPV DNA positive. Conclusion: We found a significant difference in HPV DNA test result among women above 30 years old. Co-testing of Pap and HPV DNA is needed, especially if HPV DNA type 16 and 18 were found among negative Pap results. Keywords: Cervical cancer, HPV DNA, incidence, LBC, screening   Abstrak Tujuan: Penelitian ini bertujuan melaporkan insidensi dari HPV risiko tinggi yang positif pada hasil sitologi negatif. Metode: Kami mengumpulkan 83 wanita yang menjalani liquid based cytology (LBV) dan pemeriksaan HPV DNA pada waktu yang bersamaan. Dengan menggunakan DiagCor GenoFlow Human Papilloma Virus Array Test (GenoFlow), yaitu sebuah uji HPV terbaru yang berbasis PCR dan “Flow-throug” hybridization dapat mengidentifikasi 33 subtipe HPV: 18 tipe HPV risiko tinggi seperti 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 81 dan 82. Hasil: Kami mengelompokkan subjek berdasarkan usia dibawah atau setara 30 tahun dan diatas 30 tahun (n=77). Kami menemukan perbedaansignifikan dari hasil HPV DNA dalam kelompok ini (P=0.034), dengan 19.3% memiliki HPV DNA tipe 16 dan 18 dalam kelompok usia diatas 30 tahun. Penelitian ini menunjukkan 27 perempuan (32.5%) menjalani skrining kanker serviks memiliki hasil LBC yang negatif namun menunjukkan HPV DNA positif. Kesimpulan: Terdapat perbedaan signifikan dari uji HPV DNA pada wanita usia diatas 30 tahun. Pemeriksaan bersamaan antara Pap dan HPV DNA dibutuhkan terutama ketika HPV DNA tipe 16 dan 18 ditemuka pada hasil Pap negatif. Kata kunci: HPV DNA, insidensi, kanker serviks, LBC, skrining


2000 ◽  
Vol 15 (2) ◽  
pp. 65-71
Author(s):  
Marcie Zinn ◽  
Claudia McCain ◽  
Mark Zinn

Fourteen music majors were tested using the high-risk model of threat perception (HRMTP), a biopsychosocial model designed to diagnose and guide treatment of stress-related somatic disorders. Regression analysis revealed that negative affect, social desirability, peripheral vasoconstriction, and “catastrophizing” predicted state anxiety scores after jury performance (p ≤ 0.041). A significant difference in hand temperature before and after jury performance was also found (p ≤ 0.01). Social desirability scores were inversely correlated with negative affect and catastrophizing scores (p ≤ 0.01). These results are consistent with predictions from the HRMTP, which predicts that people high in either overt or covert negative affect and catastrophizing are at greater risk for psychophysiological disorders than normals. The model also predicts that people who are high in social desirability (repressors) are likewise at risk because of inhibited pain perception. Since performance anxiety has been discussed by several authors as a psychophysiological event, implicating the role of the autonomic nervous system in the initiation and maintenance of stage fright, this model may provide a new pathway into the understanding of stage fright.


2020 ◽  
Vol 25 (3) ◽  
pp. 325-331
Author(s):  
Erkan Özmen ◽  
Ülkü Altoparlak ◽  
Muhammet Hamidullah Uyanık ◽  
Abdulkadir Gülen

Introduction: Human papillomavirus (HPV) is frequently a sexually transmitted virus and can cause cervical cancer in women. Cervical cancer is the second most common type of cancer among the developing countries. In this study, cervical HPV DNA positivity and genotype distributions were investigated in female patients living in our region and the results were compared with different studies. Materials and Methods: Between 1 July, 2017 and 1 March, 2019, 433 cervical swabs were sent to Ataturk University, Medical Faculty Hospital, Medical Microbiology Laboratory due to suspicion of HPV. Swab samples were evaluated for HPV virus using molecular (Polymerase Chain Reaction-PCR) methods. For this purpose, Xpert HPV Test (Cepheid, Inc, Sunnyvale, CA) was used to identify HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68 t in a single sample. Results: Mean age of the patients ranged from 20 to 69 years, with a mean of 39.8 years (± 10.0). Positivity was detected in 62 of the 433 patients. Mean age of the positive patients was 40.2 years (± 11.3). When the positive patients were examined in terms of HPV types, the presence of HPV 16 was observed with a rate of 25.6%, while the HPV 18/45 types were found to be 9.0% in total. When patients were evaluated according to age groups, HPV DNA positivity was highest in the 25-34 age group with 38.7%. In our statistical study, there was no significant difference in HPV DNA positivity rate between the ages of 35 and under 35 years. Conclusion: This study demonstrates the prevalence and viral genotype distribution of HPV infection in women in Erzurum region. HPV type 16 is seen with a high rate in our region.


2006 ◽  
Vol 24 (36) ◽  
pp. 5630-5636 ◽  
Author(s):  
Lisa Licitra ◽  
Federica Perrone ◽  
Paolo Bossi ◽  
Simona Suardi ◽  
Luigi Mariani ◽  
...  

Purpose Human papillomavirus (HPV) DNA tumors actively integrating the E6 and E7 oncogenes have a distinct biologic behavior resulting in a more favorable prognosis. To which extent the viral integration by itself, and/or the associated wild-type (wt) TP53 status, and/or a functional p16 contribute to prognosis is unclear. Patients and Methods To clarify how the presence of high-risk (HR) -HPV, TP53, and p16INK4a status interact with clinical outcome, we considered a retrospective series of 90 consecutive oropharyngeal cancer patients treated primarily with surgery. Results Seventeen (19%) patients showed integrated HPV 16 DNA (HPV positive), wt TP53 in all but two patients, normal p16INK4a in 15 assessable patients, and p16 expression in all 17 patients. Thirty-five patients (39%), two of whom were HPV positive, harbored TP53 mutations. p16INK4a deletion and p16 null immunophenotype occurred in 28 and 58 patients, respectively, and was similarly distributed in both patients with mutated TP53 (48% and 82%, respectively) and in patients with wt TP53 (46% and 77%, respectively). Statistical analysis showed that HPV-positive status significantly affects all investigated end points: overall survival (P = .0018), incidence of tumor relapse (P = .0371), and second tumor (P = .0152), whereas TP53 and p16INK4a status and p16 expression were not prognostic by themselves. Conclusion Our molecular and clinical results are in agreement with previous findings but provide additional information into the biologic mechanisms involved in HR-HPV oropharyngeal cancer in comparison to HPV-negative tumors. According to the reduced risk of relapse and second tumors associated with HR-HPV positivity of oropharyngeal cancer, the therapeutic strategy and follow-up procedures should be reviewed.


2013 ◽  
Vol 23 (3) ◽  
pp. 527-532 ◽  
Author(s):  
Nora Berois ◽  
Patricia De Cremoux ◽  
Daniel Mazal ◽  
Adela Sica ◽  
Mabel Cedeira ◽  
...  

ObjectivesPersistent infection with specific genotypes of human papillomaviruses (HPVs) is the main cause of invasive cervical cancer (ICC). Only a few of the various HPV types account for most of the cases worldwide, and geographical differences in their distribution are evident. Data from locally prevalent genotypes are essential in view of introduction of HPV type-specific prophylactic vaccines.MethodsIn this work, we have investigated HPV type distribution in samples of ICC cases that occurred in Uruguayan women. DNA extracted from ICC treated in Centro Hospitalario Pereira Rossell of Montevideo between 1999 and 2007 were analyzed. Search and typing were performed by polymerase chain reaction using generic GP5+/GP6+ primers and specific primers for HPV types 16, 18, 33, and 45. Positive GP5+/GP6+ samples, which were negative for all 4 high-risk HPV-specific types screened were further analyzed by sequencing.ResultsHuman papillomavirus DNA sequences were found in 163 (92.6%) of 176 cases. The most prevalent genotypes were HPV16 (67.6%) and HPV18 (8.5%) followed by HPV45 (6.8%) and HPV33 (3.4%), as single or mixed infection. Other less frequent genotypes were HPV31, HPV35, HPV39, HPV51, HPV52, HPV58, HPV66, and HPV73. The viral type could not be determined (HPV X) in 1 case (0.6%) of the HPV DNA–positive cervical cancers and double infections were found in 1.7% of the cases. The higher percentage of most aggressive HPV (16/18/45) genotypes was detected in cases diagnosed at younger than 60 years old, whereas these genotypes were less frequent in older patients.ConclusionWe conclude that HPV types 16, 18, and 45 have a very high prevalence in ICC of Uruguayan women. Results provide evidence that 16 of 18 infections are more aggressive, but most cancers could be vaccine preventable.


2005 ◽  
Vol 79 (23) ◽  
pp. 14852-14862 ◽  
Author(s):  
Jennifer C. Guess ◽  
Dennis J. McCance

ABSTRACT Infection with high-risk human papillomavirus (HPV) types, particularly types 16 and 18, contributes to 90% of cervical cancer cases. HPV infects cutaneous or mucosal epithelium, tissue that is monitored for microbial infection or damage by Langerhans cells. In lesions produced by HPV type 16, there is a reduction in numbers of immune cells, especially Langerhans cells. Langerhans precursor cells selectively express CCR6, the receptor for macrophage inflammatory protein 3α (MIP-3α), and function as potent immune responders to inflamed epithelium and initiators of the innate immune response. It has been reported that E6 and E7 of high-risk HPVs interfere with immune mediators in order to suppress the recruitment of immune cells and antiviral activities of infected cells. Here we show that, following proinflammatory stimulus, HPV-16 E6 and E7 inhibit MIP-3α transcription, resulting in suppression of the migration of immature Langerhans precursor-like cells. Interestingly, the E6 and E7 proteins from the low-risk HPV types also inhibited MIP-3α transcription. These results suggest that one mechanism by which HPV-infected cells suppress the immune response may be through the inhibition of a vital alert signal, thus contributing to the persistence of HPV infection.


2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Surapan Khunamornpong ◽  
Jongkolnee Settakorn ◽  
Kornkanok Sukpan ◽  
Suree Lekawanvijit ◽  
Narisara Katruang ◽  
...  

Objective. To evaluate the performance of high-risk human papillomavirus (HPV) DNA testing in urine samples compared to that of cervical sample testing in Northern Thailand. Methods. Paired urine and cervical samples were collected during the follow-up of women with a previous positive HPV test. HPV testing was performed using the Cobas 4800 HPV Test. Linear Array assay was used for genotyping in selected cases. Results. Paired urine and cervical samples were obtained from 168 women. Of 123 paired samples with valid results, agreement in the detection of high-risk HPV DNA was present in 106 cases (86.2%), with a kappa statistic of 0.65 (substantial agreement). Using the cervical HPV results as a reference, the sensitivity of urine HPV testing was 68.6% (24/35) and the specificity 93.2% (82/88). For the detection of histologic high-grade squamous intraepithelial lesion or worse (HSIL+), the sensitivity of urine HPV testing was 80.0% (4/5) and the specificity 78.0% (92/118). Conclusion. Although urine HPV testing had a rather low sensitivity for HPV detection, its sensitivity for histologic HSIL+ detection was high. For clinical use of urine HPV testing, standardization of specimen collection and processing techniques or application of a more sensitive test, especially in the detection of HPV52 and HPV58, is necessary.


2017 ◽  
Vol 7 (2) ◽  
pp. 91-98
Author(s):  
Mirela Dzehverovic ◽  
Anesa Ahatovic ◽  
Naris Pojskic ◽  
Naida Lojo-Kadric ◽  
Amela Pilav ◽  
...  

Introduction: Single nucleotide polymorphisms (SNPs) have lately been used for prediction of metabolic processes that may be related to obesity. The aim of our study was to examine the association of SNPs of several genes with obesity and physical activity in 18 healthy volunteers. Methods: We used buccal swabs to collect and extract DNA from 18 volunteers. Pyrosequencing was used for molecular analysis of 13 polymorphisms in 10 genes (APOA2, MTHFR, MCM6, peroxisome proliferators-activated receptor gamma, FABP2, beta-2-adrenergic receptor (ADRB)2, ADRB3, A-actinin-3, angiotensin-converting enzyme, and FUT2). The volunteers’ personal data included body mass index (BMI), dietary practice and information on daily fitness and workout routine. Association between the 13 observed gene polymorphisms and individual BMI status (normal or overweight) was analyzed. Results of the DNA analysis were used for the expert evaluation by nutritionists and physiologists to obtain optimal regulation of nutrition and exercise. The volunteers had a dietary and fitness program for 12 months which they tracked by filling in a suitable study form. Results: 14 volunteers had a moderate genetic predisposition for abdominal adipose-tissue accumulation, while 4 of them had genotypes not associated with abdominal fat tissue accumulation. A statistically significant difference was found between the value of BMI before and after the implementation of personalized training and nutrition plan within the group of overweight volunteers (paired sample t=3.382; p = 0.006; exact p = 0.015). The single-locus F-test showed no association between the gene polymorphisms and BMI values. In addition, no correlation was detected between the gene polymorphisms and amount of BMI reduction prior and after the implementation of the personalized training and nutrition plan within the overweighed group of volunteers. Conclusion: Optimal nutrition and training plan are crucial for the BMI reduction as observed in the overweighed volunteers after the 12-month personalized training and individualized nutrition plan. However, the analyzed polymorphisms were not significantly associated with the obesity in this study.


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