Evaluation of the Clinical Performance of the HPV-Risk Assay Using the VALGENT-3 Panel.

Human papillomavirus (HPV) testing is increasingly being incorporated into cervical cancer screening. The Validation of HPV Genotyping Tests (VALGENT) is a framework designed to evaluate the clinical performance of various HPV tests relative to that of the validated and accepted comparator test in a formalized and uniform manner. The aim of this study was to evaluate the clinical performance of the HPV-Risk assay with samples from the VALGENT-3 panel and to compare its performance to that of the clinically validated Hybrid Capture 2 assay (HC2). The VALGENT-3 panel comprises 1,300 consecutive samples from women participating in routine cervical cancer screening and is enriched with 300 samples from women with abnormal cytology. DNA was extracted from original ThinPrep PreservCyt medium aliquots, and HPV testing was performed using the HPV-Risk assay by investigators blind to the clinical data. HPV prevalence was analyzed, and the clinical performance of the HPV-Risk assay for the detection of cervical intraepithelial neoplasia grade 3 or worse (CIN3+) and CIN2 or worse (CIN2+) relative to the performance of HC2 was assessed. The sensitivity of the HPV-Risk assay for the detection of CIN3+ was similar to that of HC2 (relative sensitivity, 1.00; 95% confidence interval [CI], 0.95 to 1.05; P = 1.000), but the specificity of the HPV-Risk assay was significantly higher than that of HC2 (relative specificity, 1.02; 95% CI, 1.01 to 1.04; P < 0.001). For the detection of CIN2+, similar results were obtained, with the relative sensitivity being 0.98 (95% CI, 0.93 to 1.02; P = 0.257) and the relative specificity being 1.02 (95% CI, 1.01 to 1.03; P < 0.001). The performance of the HPV-Risk assay for the detection of CIN3+ and CIN2+ was noninferior to that of HC2, with all P values being ≤0.006. In conclusion, the HPV-Risk assay demonstrated noninferiority to the clinically validated HC2 by the use of samples from the VALGENT-3 panel for test validation and comparison.

Anyplex II HPV28 detection and Anyplex II HPV HR detection assays are highly concordant with other commercial assays for detection of high-risk HPV genotypes in women with high grade cervical abnormalities.

PURPOSE: to evaluate the performance of Anyplex II HPV28 and HPV HR Detection assays against the EuroArray HPV, Cobas 4800 HPV (Cobas), HPV Amplicor (Amp), Linear Array HPV (LA) and Hybrid Capture 2 (HC2) in detection of high-risk HPV (HR-HPV) from liquid-based cervical cytology samples. METHODS: cervical specimens from 404 women undergoing management of high-grade cytological abnormality were evaluated by Anyplex II HPV28 and HPV HR Detection assays for detection of HR-HPV genotypes and prediction of histologically-confirmed cervical intraepithelial neoplasia grade 2 or higher (≥CIN2). The results were compared to EuroArray, HC2, Cobas, Amp, and LA. RESULTS: specimens were evaluated from 404 women with an average age of 30 years, including 336 with a histological diagnosis of ≥ CIN2 and 68 with ≤ CIN1. Concordance of HR-HPV detection between Anyplex II HPV28 and other genotyping assays was 94.79 % (κ = 0.84; EuroArray) and 97.27 % (κ = 0.91; LA); and between Anyplex II HPV HR and other HR-HPV detection assays was 86.35 % (κ = 0.62; HC2), 96.03 % (κ = 0.87; Cobas) and 96.77 % (κ = 0.89; Amp). Using HR-HPV detection for prediction of ≥ CIN2 by Anyplex II HPV28 and HPV HR, sensitivity (90.18, 95 % CI 86.48-93.14; 90.77, 95 % CI 87.16-93.65) and specificity (both 67.16, 95 % CI 54.60-78.15) were not significantly different to the other HPV assays tested, with one exception. Both Anyplex assays had significantly higher sensitivity than HC2 (p < 0.0001), with a specificity of 96 % (p > 0.05) of HC2 in this high-risk population. CONCLUSIONS: both Anyplex II HPV detection assays were concordant with other commercial assays for HR-HPV detection, with comparable sensitivity and specificity for ≥ CIN2 detection.

Personal life and working conditions of trainees and young specialists in clinical microbiology and infectious diseases in Europe: a questionnaire survey.

The purpose of this investigation was to assess the balance between the personal and professional lives of trainees and young European specialists in clinical microbiology (CM) and infectious diseases (ID), and determine differences according to gender, country of training, workplace and specialty. The Steering Committee of the Trainee Association of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) devised a questionnaire survey consisting, beyond the demographic questions, of nine yes/no questions, 11 Likert scale self-evaluations and one open-response item on parenthood, working conditions, quality of life, alcohol consumption and burnout. This anonymous survey in English was held between April and July 2015 among European CM/ID trainees and young specialists (<3 years after training completion). Responses from 416 participants with a mean age of 32 years [standard deviation (SD) 5 years] were analysed. Females and physicians from Northern/Western Europe (NWE) benefit more from paternity/maternity leaves even during training than their counterparts. Among all respondents, only half of breastfeeding mothers enjoyed the benefit of working hours flexibility. Only two-thirds of respondents found their working environment stimulating. In comparison to colleagues from other parts of Europe, trainees and young specialists from Southern/Eastern Europe (SEE) had less frequent regular meetings with mentors/supervisors and head of departments where trainees' issues are discussed. Also, physicians from SEE were more frequently victims of workplace mobbing/bullying in comparison to those from other regions. Finally, multivariate analysis showed that female gender, SEE region and ID specialty were associated with burnout feelings. Female gender and country of work from SEE largely determine satisfactory working conditions, the possibility of parenthood leaves, amount of leisure time, mobbing experiences and burnout feelings among European CM/ID trainees and young specialists.

EUROarray human papillomavirus (HPV) assay is highly concordant with other commercial assays for detection of high-risk HPV genotypes in women with high grade cervical abnormalities.

The purpose of this study was to evaluate the performance of the EUROIMMUN EUROArray HPV genotyping assay against the Roche Cobas 4800, Roche HPV Amplicor, Roche Linear Array and Qiagen Hybrid Capture 2 assays in the detection of high-risk HPV (HR-HPV) from liquid based cervical cytology samples collected from women undergoing follow-up for abnormal cervical cytology results. Cervical specimens from 404 women undergoing management of high-grade cytological abnormality were evaluated by EUROarray HPV for detection of HR-HPV genotypes and prediction of histologically-confirmed cervical intraepithelial neoplasia grade 2 or higher (≥CIN2). The results were compared to Hybrid Capture 2, Cobas 4800 HPV, Amplicor and Linear Array HPV. Positivity for 14 HR-HPV types was 80.0 % for EUROarray (95 % CI; 75.7-83.8 %). Agreement (κ, 95 % CI) between the EUROarray and other HPV tests for detection of HR-HPV was good to very good [Hybrid Capture κ = 0.62 (0.54-0.71); Cobas κ = 0.81 (0.74-0.88); Amplicor κ = 0.68 (0.60-0.77); Linear Array κ = 0.77 (0.70-0.85)]. For detection of HR-HPV, agreement with EUROarray was 87.90 % (Hybrid Capture), 93.58 % (Cobas), 92.84 % (Amplicor) and 92.59 % (Linear Array). Detection of HR-HPV was not significantly different between EUROarray and any other test (p < 0.001). EUROarray was concordant with other assays evaluated for detection of high-risk HPV and showed sensitivity and specificity for detection of ≥ CIN2 of 86 % and 71 %, respectively.

Primary resistance to integrase strand-transfer inhibitors in Europe.

OBJECTIVES: The objective of this study was to define the natural genotypic variation of the HIV-1 integrase gene across Europe for epidemiological surveillance of integrase strand-transfer inhibitor (InSTI) resistance. METHODS: This was a multicentre, cross-sectional study within the European SPREAD HIV resistance surveillance programme. A representative set of 300 samples was selected from 1950 naive HIV-positive subjects newly diagnosed in 2006-07. The prevalence of InSTI resistance was evaluated using quality-controlled baseline population sequencing of integrase. Signature raltegravir, elvitegravir and dolutegravir resistance mutations were defined according to the IAS-USA 2014 list. In addition, all integrase substitutions relative to HXB2 were identified, including those with a Stanford HIVdb score ≥ 10 to at least one InSTI. To rule out circulation of minority InSTI-resistant HIV, 65 samples were selected for 454 integrase sequencing. RESULTS: For the population sequencing analysis, 278 samples were retrieved and successfully analysed. No signature resistance mutations to any of the InSTIs were detected. Eleven (4%) subjects had mutations at resistance-associated positions with an HIVdb score ≥ 10. Of the 56 samples successfully analysed with 454 sequencing, no InSTI signature mutations were detected, whereas integrase substitutions with an HIVdb score ≥ 10 were found in 8 (14.3%) individuals. CONCLUSIONS: No signature InSTI-resistant variants were circulating in Europe before the introduction of InSTIs. However, polymorphisms contributing to InSTI resistance were not rare. As InSTI use becomes more widespread, continuous surveillance of primary InSTI resistance is warranted. These data will be key to modelling the kinetics of InSTI resistance transmission in Europe in the coming years.

Hepatic expression of miR-122, miR-126, miR-136 and miR-181a and their correlation to histopathological and clinical characteristics of patients with hepatitis C.

It has been shown for hepatitis C virus (HCV) infection that host miRNAs contribute to the replication of the viral RNA genome. However, the clinical impact of these and many other cellular miRNAs on HCV in humans is still largely unclear. We therefore analysed the expression of miR-122, miR-126, miR-181a and miR-136 in HCV-infected patients. The study included liver biopsies of 65 patients infected with HCV of different genotypes (gt 1, gt 1a, gt 1b, gt 3 and gt 4) and nine noninfected individuals. Expression analysis of miRNAs was performed by qPCR, and they were analysed for differences between patient gender and age, genotypes, stage of fibrosis, grade of inflammation, serum level of liver enzymes, serum viral load, the presence of steatosis and mode of transmission. Different target prediction algorithms were used to search for targets of analyzed miRNAs. Statistical analysis revealed significant up-regulation of miR-136 and down-regulation of miR-126 and miR-181a in patients infected with HCV of different genotypes compared with noninfected individuals. The same expression pattern was observed in different stages and grades of liver disease. miR-122 was up-regulated in women relative to men and associated to portal inflammation, miR-122 and miR-126 correlated with serum HCV load and miR-136 and miR-122 correlated with the presence of steatosis. miR-126 and miR-136 were differentially expressed between different modes of HCV transmission. There were approximately 2000 different targets predicted for all four miRNAs and each of the analyzed miRNAs could be involved in more than a 100 different biochemical pathways. miR-122, miR-126, miR-136 and miR-181a have been shown to be involved in HCV infection with different genotypes. Their expression has been associated with the gender, stage and grade of liver disease, mode of transmission, serum HCV load and the presence of steatosis. Numerous target genes and biochemical pathways are predicted for each of the analyzed miRNAs. All these results suggest their role in HCV-infected liver disease.

Hepatitis C virus infection among pregnant women in Slovenia: study on 31,849 samples obtained in four screening rounds during 1999, 2003, 2009 and 2013.

The majority of people infected with hepatitis C virus (HCV) are unaware of their infection. Assessment of the prevalence of HCV infection in the general population and in key populations at increased risk is needed for evidence-based testing policies. Our objectives were to estimate the prevalence of antibodies to HCV (anti-HCV), the prevalence of HCV viraemia (HCV RNA), and to describe HCV genotype distribution among pregnant women in Slovenia. Unlinked anonymous testing was performed on residual sera obtained from 31,849 pregnant women for routine syphilis screening during 1999, 2003, 2009, and 2013. Anti-HCV reactive specimens were tested for HCV RNA and HCV genotypes were determined. Annual prevalence of anti-HCV ranged between 0.09% (95% confidence interval (CI): 0.03­0.18) in 2009 and 0.21% (95% CI: 0.12­0.34) in 2003 and HCV RNA positivity between 0.06% (95% CI: 0.02­0.14) in 2009 and 0.14% (95% CI: 0.07­0.25) in 2003. We observed no statistically significant differences in anti-HCV or HCV RNA prevalence between age groups (<20, 20­29 and ≥30 years) in any year and no trend in time. Of 29 HCV active infections, 19 were with genotype 1 and 10 with genotype 3. HCV infection among pregnant women was rare suggesting a low burden in the Slovenian general population. Antenatal screening for HCV in Slovenia could not be recommended.

Validation of the clinical performance and reproducibility of the NeuMoDx HPV assay self-sample workflow.

BACKGROUND: Human papillomavirus (HPV) testing on self-samples is a valid tool for cervical cancer screening. HPV self-sample workflows need to be clinically validated to ensure safe use in screening. OBJECTIVE: This study evaluated the fully automated NeuMoDx HPV Assay self-sample workflow that is compiled of the NeuMoDx HPV assay and the NeuMoDx 96/288 Molecular Systems, for clinical performance and reproducibility on Evalyn Brush-collected self-samples. METHODS: The clinical performance of the NeuMoDx HPV Assay self-sample workflow for cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and CIN3+ was evaluated on 987 self-samples obtained from women attending national organized HPV-based cervical cancer screening by a noninferiority analysis relative to reference workflows using either HPV-Risk Assay or high-risk HPV GP5+/6+-PCR. Intra- and inter-laboratory reproducibility of the NeuMoDx HPV Assay self-sample workflow using both NeuMoDx 96 and 288 Molecular Systems was assessed on 520 self-samples in three laboratories. RESULTS: The clinical sensitivity and specificity of the NeuMoDx HPV Assay self-sample workflow for the detection of CIN2+ and CIN3+ were found to be non-inferior to the reference workflows using either HPV-Risk Assay or high-risk HPV GP5+/6+-PCR, with all p-values <0.034. The NeuMoDx HPV Assay self-sample workflow exhibited an intra-laboratory reproducibility of 94.4 % (95 %CI:92.5-96.1 %) with kappa value 0.86 (95 %CI:0.81-0.91). Inter-laboratory agreement was high (all ≥93.4 % and all kappa values ≥0.83). CONCLUSIONS: The NeuMoDx HPV Assay self-sample workflow demonstrated high clinical accuracy for CIN2+/3+ and high reproducibility. The NeuMoDx HPV Assay self-sample workflow can be considered suitable for cervical cancer screening purposes.

Clinical validation of the Abbott RealTime High Risk HPV assay according to the guidelines for human papillomavirus DNA test requirements for cervical screening.

This study showed that the Abbott RealTime High Risk HPV assay fulfilled cross-sectional clinical equivalence and reproducibility criteria of international consensus guidelines, which indicates that this assay can be considered clinically validated for cervical cancer screening purposes.

Primary HPV-based cervical cancer screening in Europe: implementation status, challenges, and future plans.

BACKGROUND: Cytology-based screening has been a cornerstone of cervical cancer prevention for decades. Following extensive evidence demonstrating higher sensitivity and accuracy, lower variability and better reproducibility of human papillomavirus (HPV)-based screening compared with conventional or liquid-based cytology, recent European guidelines strongly recommend primary HPV-based screening over standard cytology-based screening. In addition, HPV-based screening offers the possibility of self-sampling and makes possible longer screening intervals in women with negative screening results. OBJECTIVES: We summarize the current status of implementation of HPV-based screening in Europe, describe the real-life experience and challenges from countries already performing HPV-based screening, and briefly review immediate and long-term plans for screening implementation in selected European countries. SOURCES: Data were obtained from peer-reviewed literature, personal communication with experts and authorities involved in formulating national recommendations and practical guidelines, and relevant national websites. CONTENT: As of July 2019, the Netherlands and Turkey are the only European countries with fully implemented national HPV-based cervical cancer screening. Italy, Sweden and Finland have already implemented HPV-based screening in several regions, and several other countries are at various stages of implementation. Some countries are considering transitioning from cytology-based to HPV-based screening, but are struggling with the suboptimal performance of current population-based programmes. Implementation of HPV-based screening has resulted in higher colposcopy referral rates, but also higher detection rates of CIN3+ lesions and cervical cancers requiring immediate treatment. Cytology is mostly used as a triage test, although other strategies are under consideration in some countries. IMPLICATIONS: HPV-based screening is best suited in organized population-based screening settings. In 2019, cervical cancer screening policies across Europe vary greatly. Experience in countries with national and regional HPV-based screening already implemented is generally very positive. Urgent action is needed in many European countries, especially those with suboptimal opportunistic cytology-based cervical cancer screening.

Sniffing animals as a diagnostic tool in infectious diseases.

BACKGROUND: Scents and odours characterize some microbes when grown in the laboratory, and experienced clinicians can diagnose patients with some infectious diseases based on their smell. Animal sniffing is an innate behaviour, and animals' olfactory acuity is used for detecting people, weapons, bombs, narcotics and food. OBJECTIVES: We briefly summarized current knowledge regarding the use of sniffing animals to diagnose some infectious diseases and the potential use of scent-based diagnostic instruments in microbiology. SOURCES: Information was sought through PubMed and extracted from peer-reviewed literature published between January 2000 and September 2019 and from reliable online news. The search terms 'odour', 'scent', 'bacteria', 'diagnostics', 'tuberculosis', 'malaria' and 'volatile compounds' were used. CONTENT: Four major areas of using sniffing animals are summarized. Dogs have been used to reliably detect stool associated with toxigenic Clostridioides difficile and for surveillance. Dogs showed high sensitivity and moderate specificity for detecting urinary tract infections in comparison to culture, especially for Escherichia coli. African giant pouched rats showed superiority for diagnosing tuberculosis over microscopy, but inferiority to culture/molecular methods. Several approaches for detecting malaria by analysing host skin odour or exhaled breath have been explored successfully. Some microbial infections produce specific volatile organic compounds (VOCs), which can be analysed by spectrometry, metabolomics or other analytical approaches to replace animal sniffing. IMPLICATIONS: The results of sniffing animal studies are fascinating, and animal sniffing can provide intermediate diagnostic solutions for some infectious diseases. Lack of reproducibility, and cost of animal training and housing are major drawbacks for wider implementation of sniffing animals. The ultimate goal is to understand the biological background of this animal ability and to characterize the specific VOCs that animals are recognizing. VOC identification, improvement of odour sampling methods and development of point-of-care instruments could allow implementation of scent-based tests for major human pathogens.

Use of drones in clinical microbiology and infectious diseases: current status, challenges and barriers.

BACKGROUND: Drones or unmanned aerial vehicles are autonomous or remotely controlled multipurpose aerial vehicles driven by aerodynamic forces and capable of carrying a payload. Whereas initially used exclusively for military purposes, the use of drones has gradually spread into other areas. Given their great flexibility and favourable costs, the use of drones has also been piloted in various healthcare settings. OBJECTIVES: We briefly summarize current knowledge regarding the use of drones in healthcare, focusing on infectious diseases and/or microbiology when applicable. SOURCES: Information was sought through PubMed and extracted from peer-reviewed literature published between January 2010 and August 2019 and from reliable online news sources. The search terms 'drones', 'unmanned aerial vehicles', 'microbiology' and 'medicine' were used. CONTENT: Peer-reviewed literature on the use of drones in healthcare has steadily increased in recent years. Drones have been successfully evaluated in various pilot programmes and are already implemented in some settings for transporting samples and delivering blood, vaccines, medicines, organs, life-saving medical supplies and equipment. In addition, a promising proof-of-concept 'lab-on-a-drone' was recently presented, as well as several pilot studies showing the benefits of drone use in surveillance and epidemiology of infectious diseases. IMPLICATIONS: The potential for drone use in clinical microbiology, infectious diseases and epidemiology is vast. Drones may help to increase access to healthcare for individuals that might otherwise not benefit from appropriate care due to remoteness and lack of infrastructure or funds. However, factors such as national airspace legislation and legal medical issues, differences in topography and climates, cost-effectiveness, and community attitudes and acceptance in different cultures and societies currently impede the widespread use of drones. Significant cost savings compared with ground transportation, speed and convenience of delivery, and the booming drone sector will probably drive drone implementation in various areas of medicine in the next 5 years.

Portable molecular diagnostic instruments in microbiology: current status.

BACKGROUND: Mobile microbiology is an evolving concept that has the potential to reduce morbidity and mortality associated with infectious diseases on a global level. Molecular methods used in the context of mobile microbiology ensure rapid and accurate aetiological diagnostics and allow timely initiation of clinical care. The great majority of published data regarding molecular diagnostics in mobile laboratories have focused on emerging viral infections and using laboratory-developed assays. Use of clinically validated and commercially available molecular diagnostic instruments in routine diagnostics of infectious diseases in mobile laboratories has received only limited attention in the field. OBJECTIVES: This review summarizes the suitability of a range of portable diagnostic molecular instruments for application in mobile laboratories by taking into account the instruments' analytical concepts, technical features and environmental requirements, as well as results of major validation studies. SOURCES: Data on technical features of selected portable instruments were mainly extracted from manufacturers' websites. Information on validation studies of various molecular assays developed for the selected instruments was extracted from peer-reviewed publications searched for through PubMed. CONTENT: Eight portable diagnostic molecular instruments (Alere q, GeneXpert Edge, GeneXpert Omni, Genedrive, PanNAT, Revogene, cobas Liat and ID Now) that are commercially available or in the launching stage are presented and evaluated in the context of the mobile microbiology concept, with particular emphasis on technical features and environmental requirements. Both the cobas Liat and the Alere i assays have been extensively validated in a variety of studies carried out in both adult and paediatric patients from various settings (ranging from primary care to emergency care departments in tertiary centres). Most studies showed comparable performance of cobas Liat and Alere i molecular assays with the standard-of-care in vitro diagnostics molecular assays routinely performed in dedicated/centralized molecular diagnostics laboratories. In addition, acceptable performance of Alere q and Genedrive instruments has been shown in implementations studies for early infant diagnosis of children born to human immunodeficiency virus-positive mothers and detection of hepatitis C virus RNA, respectively. Additional validation studies on existing (GeneXpert Edge, PanNAT, Revogene) and emerging (GeneXpert Omni) technologies are warranted. IMPLICATIONS: Several portable molecular diagnostic platforms reviewed are suitable for mobile microbiology applications. Further development in this field should be directed toward providing a broader range of assays per instrument, multiplexing, reducing the frequency of invalid results, and price cutting.

Smartphones as mobile microbiological laboratories.

BACKGROUND: Point-of-care (POC) tests provide an alternative to traditional laboratory-based diagnostics due to reduced turnaround times, portability and no need for highly trained laboratory staff. Smartphones can be integrated into POC platforms because of their multifunctionality, enabled by high-quality digital cameras, computer processors, touchscreen interface and wireless data transfer. It is predicted that by 2020 about 80% of the world population will use smartphones. OBJECTIVES: This review summarizes the current state of the art regarding smartphones as part of a mobile microbiological laboratory. SOURCES: Selected peer-reviewed publications on smartphone-based microbiological testing published between January 2015 and August 2019. CONTENT: Smartphones can be used as instrumental interfaces, dongles, microscopes or test result readers (brightfield, colorimetric and fluorescent measurements), or combined with amplification methods such as loop-mediated isothermal amplification (LAMP) tests in portable POC test platforms. Smartphone-based tests offer opportunities for microbiological diagnostics in remote areas and both resource-limited and resource-rich settings. Wireless connectivity may facilitate epidemiological studies and creation of spatiotemporal disease prevalence maps. However, the current analytical performance of many smartphone-based POC tests must be improved and carefully validated in clinical settings by comparison with current diagnostic standards. IMPLICATIONS: Recent developments in smartphone-based POC tests for infectious diseases are promising, as evidenced by results from many proof-of-concept studies. Further progress will foster large-scale implementation of smartphone-based POC as mobile microbiological laboratories in the near future.

Commercially available molecular tests for human papillomaviruses: a global overview.

BACKGROUND: Molecular tests for detection of human papillomaviruses (HPVs) play a crucial role in the prevention of cervical cancer, including recently announced elimination efforts. HPV testing is a recommended approach for cervical cancer screening of women over 30 and for management of those with precancerous cervical lesions. In addition, they are widely used in epidemiological studies, HPV surveillance and vaccination impact monitoring. OBJECTIVES: The aim was to provide an updated 2020 inventory of commercial molecular HPV tests available on the market. SOURCES: Data were retrieved from internal files, and a detailed search using Medline/Pubmed, Web of Science, Scopus, Google Scholar, Google and Bing, without language or period restrictions, was performed in September 2019 and again in January 2020. CONTENT: We identified 254 distinct commercial HPV tests and at least 425 test variants available on the global market in 2020, which represents a 31% and 235% increase in the number of distinct tests and variants, respectively, compared with the previous inventory performed in 2015. Although the proportion of commercially available HPV tests with at least one peer-reviewed publication has increased over the past decade, 60% of the HPV tests on the global market are still without a single peer-reviewed publication. Furthermore, 82% of tests lack any published analytical and/or clinical evaluation, and over 90% are not evaluated in line with consensus requirements that ensure safe use in clinical settings. IMPLICATIONS: Significant challenges and scope for improvement still exist for both the HPV scientific community and the manufacturers of HPV tests. The latter must put more effort into validating their products, in agreement with standardized procedures, including all steps of HPV testing and various clinical specimens. High throughput capacity and point-of-care HPV tests are needed, both with affordable prices.

Low prevalence of active COVID-19 in Slovenia: a nationwide population study of a probability-based sample.

OBJECTIVES: Accurate population-level assessment of the coronavirus disease 2019 (COVID-19) burden is fundamental for navigating the path forward during the ongoing pandemic, but current knowledge is scant. We conducted the first nationwide population study using a probability-based sample to assess active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, combined with a longitudinal follow-up of the entire cohort over the next 6 months. Baseline SARS-CoV-2 RNA testing results and the first 3-week follow-up results are presented. METHODS: A probability-based sample of the Slovenian population comprising data from 2.1 million people was selected from the Central Population Register (n = 3000). SARS-CoV-2 RNA was detected in nasopharyngeal samples using the cobas 6800 SARS-CoV-2 assay. Each participant filled in a detailed baseline questionnaire with basic sociodemographic data and detailed medical history compatible with COVID-19. After 3 weeks, participants were interviewed for the presence of COVID-19-compatible clinical symptoms and signs, including in household members, and offered immediate testing for SARS-CoV-2 RNA if indicated. RESULTS: A total of 1368 individuals (46%) consented to participate and completed the questionnaire. Two of 1366 participants tested positive for SARS-CoV-2 RNA (prevalence 0.15%; posterior mean 0.18%, 95% Bayesian confidence interval 0.03-0.47; 95% highest density region (HDR) 0.01-0.41). No newly diagnosed infections occurred in the cohort during the first 3-week follow-up round. CONCLUSIONS: The low prevalence of active COVID-19 infections found in this study accurately predicted the dynamics of the epidemic in Slovenia over the subsequent month. Properly designed and timely executed studies using probability-based samples combined with routine target-testing figures provide reliable data that can be used to make informed decisions on relaxing or strengthening disease mitigation strategies.

2008 European Guideline on HIV testing.

Testing for HIV is one of the cornerstones in the combat against HIV infection. The 2008 European Guideline on HIV Testing provides advice on testing for HIV infection in individuals aged 16 years and older who have sought evaluation and treatment at sexually transmitted infection services for dermatovenereology clinics across Europe. Its aim is to provide practical guidance to clinicians in these settings who undertake HIV testing and suggest appropriate standards for the audit of service provision.

Comparison of paired cervical scrape and tumor tissue samples for detection of human papillomaviruses in patients with cervical cancer.

PURPOSE OF INVESTIGATION: To compare the detection and distribution of HPV genotypes in paired cervical scrape samples and tumor tissue samples in patients with cervical cancer. METHODS: Forty cervical scrape samples and 40 paired archival or fresh frozen tissue samples were collected from women with cervical cancer. Polymerase chain reaction with GP5+ and GP6+ primers was performed in all samples for HPV DNA detection. All GP5+/GP6+ negative samples were additionally tested using INNO-LiPA HPV Genotyping Extra Test. RESULTS: Overall, 39/40 (97.5%) of CC samples were HPV DNA positive. HPV 16 was found in 24/40 samples, HPV 18 in 5/40 samples. A co-infection with two different HPV genotypes was identified in one cervical scrape specimen, while in tissue samples only single infections were detected. Overall agreement between paired samples was 98.75%. CONCLUSION: The present study has shown that cervical scrape samples are equally useful for HPV genotype determination as tumor tissue samples in patients with cervical cancer. They can be used as accurate clinical samples for detection of HPV genotype causing cervical cancer or for epidemiological molecular studies.

Human telomerase gene amplification and high-risk human papillomavirus infection in women with cervical intra-epithelial neoplasia.

This study was designed to investigate whether a correlation exists between amplification of the human telomerase gene (human telomerase RNA component [TERC]) and high-risk human papillomavirus (HR-HPV) infection in 101 women with cervical intra-epithelial neoplasia (CIN). Eight patients (7.9%) had CIN 1, 24 (23.8%) had CIN 2 and 69 (68.3%) had CIN 3. TERC was amplified in 31.7% of all CIN patients. The difference in frequency of TERC amplification between patients with low-grade CIN (CIN 1) and those with high-grade CIN (CIN 2 and CIN 3) was not significant. HR-HPV infection was detected in 88.1% of all CIN cases and was significantly more frequent in patients with CIN 2 and CIN 3 than in patients with CIN 1. There was no significant difference in the frequency of HR-HPV infection between groups of patients with and without TERC amplification. In conclusion, this study found no correlation between TERC amplification and HR-HPV infection in patients with CIN.

Pre-vaccination distribution of human papillomavirus (HPV) genotypes in women with cervical intraepithelial neoplasia grade 3 (CIN 3) lesions in Slovenia.

BACKGROUND: High-risk human papillomaviruses (HPV) are the main etiological factor of cervical cancer. Cervical intraepithelial neoplasia grade 3 (CIN 3) is the latest pre-invasive stage of cervical cancer, with an approximately 20% progression rate to invasive cervical carcinoma. OBJECTIVE: To establish the pre-vaccination distribution of HPV genotypes in Slovenian women with CIN 3 lesions, in order to assess the potential benefit of prophylactic HPV vaccination in Slovenia, and to provide baseline data for monitoring the potential replacement of HPV genotypes under selective pressure of HPV vaccines. METHODS AND RESULTS: A total of 261 cervical swabs collected from women with histologically confirmed CIN 3 lesions were analyzed using four genotyping methods: the Abbott RealTime High Risk HPV Assay, the Innogenetics INNO-LiPA HPV Genotyping Extra Test, and the in-house PGMY09/11, and CPI/CPIIg polymerase chain reaction (PCR) and sequencing. Of 261 samples, 253 (96.9%) were HPV positive. The most common HPV genotype in CIN 3 lesions in the Slovenian samples was HPV-16 (59.0%), followed by HPV-31 (7.5%), HPV-33 (7.1%), HPV-58 (5.0%), and HPV-51 (4.0%). The presence of more than one HPV genotype was detected in 49/253 (19.4%) samples. Together, HPV-16 and HPV- 18 accounted for 67.4% of CIN 3 lesions in this Slovenian population. CONCLUSION: The high proportion of CIN 3 lesions caused by HPV-16 and HPV-18 should further support the recent decision to include the prophylactic vaccination against HPV in the national vaccination program in Slovenia.