A clinical utility evaluation of dual HIV/Syphilis point-of-care tests in non-clinical settings for screening for HIV and syphilis in men who have sex with men.

BACKGROUND: Dual point-of-care tests (POCTs) for the simultaneous detection of antibodies to HIV and syphilis have been developed. Since community-based organisations (CBO) are effective providers of HIV and syphilis testing among men who have sex with men (MSM), evaluation of the utility of these dual tests at CBO testing services is a high priority. The aim of this study is to determine the feasibility of performing dual HIV-syphilis POCT testing among both users and providers at these non-clinical sites. METHODS: This evaluation assessed the utility of two lateral flow immunochromatographic antibody technologies for dual screening for HIV/syphilis among MSM seeking testing in four CBO testing services in Spain, Slovenia, Latvia, and Ukraine. The study's conceptual framework divides the concept of feasibility into two inter-related domains, acceptability, and usability and further breaks it down into six subdomains: learnability, willingness, suitability, satisfaction, efficacy, and effectiveness. The feasibility analysis was performed by calculating the median score in 3 stages (for individual questions, subdomains, and domains), using a summated scores method. RESULTS: The final sample included 844 participants, 60 of which were found to be HIV test positive (7.1%) and 61 (7.2%) positive on testing for syphilis. There was a small difference (1.1%) when comparing the results of the two dual POCTs under evaluation to the tests routinely used at each site. The inter-rater agreement showed a high concordance between two independent readings. The analysis of the feasibility for the users of the services indicated good satisfaction, suitability, and willingness. In addition, among 18 providers the total mean score showed good acceptability and usability, good willingness, easy learnability, high suitability, and good efficacy, but lower satisfaction and effectiveness. The operational characteristics of both dual study POCTs were well evaluated by providers. CONCLUSIONS: The introduction of dual HIV and syphilis  POCTs in CBO testing services for screening of MSM is feasible, with a high acceptability and usability both for users and providers. Implementation of dual POCTs for HIV and syphilis in CBO testing services is an opportunity for scaling up integrated HIV/syphilis testing for MSM.

Microbiome-derived cobalamin and succinyl-CoA as biomarkers for improved screening of anal cancer.

Human papillomavirus can cause preinvasive, high-grade squamous intraepithelial lesions (HSILs) as precursors to cancer in the anogenital area, and the microbiome is suggested to be a contributing factor. Men who have sex with men (MSM) living with human immunodeficiency virus (HIV) have a high risk of anal cancer, but current screening strategies for HSIL detection lack specificity. Here, we investigated the anal microbiome to improve HSIL screening. We enrolled participants living with HIV, divided into a discovery (n = 167) and validation cohort (n = 46), and who were predominantly (93.9%) cisgender MSM undergoing HSIL screening with high-resolution anoscopy and anal biopsies. We identified no microbiome composition signatures associated with HSILs, but elevated levels of microbiome-encoded proteins producing succinyl coenzyme A and cobalamin were significantly associated with HSILs in both cohorts. Measurement of these candidate biomarkers alone in anal cytobrushes outperformed anal cytology as a diagnostic indicator for HSILs, increasing the sensitivity from 91.2% to 96.6%, the specificity from 34.1% to 81.8%, and reclassifying 82% of false-positive results as true negatives. We propose that these two microbiome-derived biomarkers may improve the current strategy of anal cancer screening.

Metagenomic Mining for Esterases in the Microbial Community of Los Rueldos Acid Mine Drainage Formation.

Acid mine drainage (AMD) systems are extremely acidic and are metal-rich formations inhabited by relatively low-complexity communities of acidophiles whose enzymes remain mostly uncharacterized. Indeed, enzymes from only a few AMD sites have been studied. The low number of available cultured representatives and genome sequences of acidophiles inhabiting AMDs makes it difficult to assess the potential of these environments for enzyme bioprospecting. In this study, using naïve and in silico metagenomic approaches, we retrieved 16 esterases from the α/ß-hydrolase fold superfamily with the closest match from uncultured acidophilic Acidobacteria, Actinobacteria (Acidithrix, Acidimicrobium, and Ferrimicrobium), Acidiphilium, and other Proteobacteria inhabiting the Los Rueldos site, which is a unique AMD formation in northwestern Spain with a pH of ∼2. Within this set, only two polypeptides showed high homology (99.4%), while for the rest, the pairwise identities ranged between 4 and 44.9%, suggesting that the diversity of active polypeptides was dominated not by a particular type of protein or highly similar clusters of proteins, but by diverse non-redundant sequences. The enzymes exhibited amino acid sequence identities ranging from 39 to 99% relative to homologous proteins in public databases, including those from other AMDs, thus indicating the potential novelty of proteins associated with a specialized acidophilic community. Ten of the 16 hydrolases were successfully expressed in Escherichia coli. The pH for optimal activity ranged from 7.0 to 9.0, with the enzymes retaining 33-68% of their activities at pH 5.5, which was consistent with the relative frequencies of acid residues (from 54 to 67%). The enzymes were the most active at 30-65°C, retaining 20-61% of their activity under the thermal conditions characterizing Los Rueldos (13.8 ± 0.6°C). The analysis of the substrate specificity revealed the capacity of six hydrolases to efficiently degrade (up to 1,652 ± 75 U/g at pH 8.0 and 30°C) acrylic- and terephthalic-like [including bis(2-hydroxyethyl)-terephthalate, BHET] esters, and these enzymes could potentially be of use for developing plastic degradation strategies yet to be explored. Our assessment uncovers the novelty and potential biotechnological interest of enzymes present in the microbial populations that inhibit the Los Rueldos AMD system.

A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions.

Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR2 E2 , with efficient native transaminase (kcat : 69.49±1.77 min-1 ) and artificial esterase (kcat : 3908-0.41 min-1 ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR2 E2 (pHopt : 8.0-9.5; Topt : 60-65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the ß-keto ester into the ß-keto acid at the hydrolytic site and subsequently into the ß-amino acid (e.e. >99 %) at the transaminase site. The catalytic power of the TR2 E2 PluriZyme was proven with a set of ß-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.

Integration of community-based testing data into national HIV surveillance in Poland, Serbia and Slovakia within the framework of INTEGRATE project.

BACKGROUND: Community-based voluntary counselling and testing contributes to early HIV diagnoses among key populations. Testing data from such decentralized services is however often not standardized nor linked to national surveillance systems. This study aimed to support the integration of community testing data into respective national surveillance and monitoring and evaluation systems for those infections. We present results from three national pilots, focused on improved data collection and transfer. METHODS:  Within the Joint Action INTEGRATE different pilot activities were planned and implemented according to the local context. In Slovakia, standardised data collection tools were implemented in three community testing services. The data generated was used to calculate the proposed indicators. In Poland, positive test results from the community testing database were linked to the national case-based surveillance database using confirmatory test number, to improve the completeness of behavioural data in the national database. In Serbia, voluntary counselling and testing forms were improved enabling identification of community-based testing. A system to generate unique client identifiers was initiated in the National registry of HIV cases to monitor linkage to care. RESULTS: All three sites were able to estimate most of the agreed indicators. In Slovakia during the study period 675 people were tested for HIV, 410 for hepatitis C and 457 for syphilis, with reactivity rates of 0.4, 2.5 and 1.8%, respectively. For HIV, 66.7% of reactive cases were confirmed and linked to care. In Poland, 28.9% of the community testing sites' records were linked to the national surveillance database (and accounted for 14.3% of all new diagnoses registered here during 2017-2018). Reactivity rate ranged between 1.9% and 2.1%. In Serbia, 80 persons were tested at community sites, from which two had a reactive HIV test result. By linking unique client identifiers from voluntary counselling and testing and National Registry of HIV cases databases, linkage to care within a two-month period was observed for one of two people with reactive HIV test result. CONCLUSIONS:  Pilot activities in the three countries demonstrate that integration of community-based testing data into surveillance systems is feasible and can help improve national surveillance data by providing key information.

Opinions Towards Key Operational Aspects for the Implementation of HIV Self-Testing in Spain: A Comparison between Stakeholders and Potential Users.

We assessed previous knowledge about the existence of HIV self-testing of stakeholders in Spain, as well as their personal position towards this methodology. We also assessed their views on potential users' (PU) opinions towards several key operational aspects surrounding self-testing, and compared them to those expressed by a sample of PU comprised of men who have sex with men. In 2017, we recruited three types of stakeholders: public health professionals and policy makers (PHPPM) (n = 33), clinical providers (n = 290) and community-based/non-governmental organization (CBO/NGO) workers (n = 55). Data on PU (n = 3537) were collected in 2016. Previous knowledge about the existence of self-testing was higher in stakeholders than in PU, but being in favor was less frequent. PUs' willingness to pay 25-30 euros for a self-test was higher than that which stakeholders considered. According to clinical providers and PHPPM, pharmacies would be PUs' preferred place to obtain a self-test, which was in line with PUs' actual choice. CBO/NGO workers on the other hand thought it would be CBO/NGOs. PHPPM and clinical providers considered primary care as PUs' preferred setting to confirm a reactive self-test and CBO/NGO chose CBO/NGOs, but PUs preferred an HIV/STI testing service or clinic. Stakeholders' opinions significantly differed from those of PUs. This divergence needs to be brought up to stakeholders as it could vary their position towards self-testing as well as the actions taken in the implementation of a testing option with the potential of increasing testing frequency.

[HIV screening in leisure spaces in Barcelona and Sitges in men who have sex with men]. / Cribado del VIH en espacios de ocio en Barcelona y Sitges en hombres que tienen sexo con hombres.

Description of the implementation of the field work of a pilot intervention whose objective was to promote the early diagnosis of HIV infection by offering a screening test for HIV in gay venues and cruising areas (outdoor sexual contact areas) frequented by men who have sex with men (MSM) in the city of Barcelona and Sitges, and consulting the result through the project website. The pilot intervention was viable and counted with the acceptability of the target group.

Hepatitis C Screening in Community-Based Voluntary Counselling and Testing Services in Europe: An Observational Study from the COBATEST Network 2014-2018.

The COBATEST Network links community-based voluntary counselling and testing (CBVCT) services in the European region and collects testing data using standardised data collection tools. This study aims to describe the population being screened for anti-HCV antibodies in the COBATEST Network and identify risk factors associated with a reactive HCV screening test result in the period 2014-2018. Clients aged > 16 screened for HCV in the period 2014-2018 at one of the Network's CBVCT services were included in the study. In the 5 year period, 7426 clients were screened for HCV in 22 centres in 10 countries and anti-HCV antibodies were detected in 113 (1.5%). The majority of people screened were aged 25-44, men who have sex with men (MSM), not HIV+ , not reporting a history of injecting drug use or sex work. Detection of anti-HCV antibodies was associated with being HIV + MSM (aOR 9.1, 95% CI 3.8; 21.8 compared to HIV-clients) and being a person who injects drugs (PWID, aOR 28.1, 95% CI 17.6; 45.0, compared to people with no history of injecting drug use). This study demonstrates that HIV-MSM with no history of injection drug use are using CBVCT services for HCV screening, but reactive screening test is associated with being HIV+ or PWID. The integration of HCV screening into the CBVCT service model may widen access to testing for populations that may otherwise not be tested.

[Cobatest network: users' characteristics of community-based voluntary,counselling and testing centres in Spain.] / Red COBATEST: Características de los usuarios de los centros comunitarios de diagnóstico y orientación para el VIH en España.

OBJECTIVE: The COBATEST network (European network of community-based voluntary, counselling and testing centers) allows standardized collection of information to monitor HIV testing at these centers. The objective was to describe the performance of the Spanish centers of the COBATEST network and the epidemiological profile of its users, and to analyze the factors associated with having an earlier HIV test. METHODS: Descriptive analysis of the data collected in the 19 Spanish centers that used the common information collection tools during the year 2015. A multivariate logistic regression model was used to identify factors associated with having an earlier test. RESULTS: A total of 6,077 HIV rapid tests were performed and 1.8% were reactive (3.0% MSM, 3.1% SW, 2.3% PWID, 0.6% heterosexual). 82.1% of the reactives were referred to perform a confirmatory test and 94 (97.9%) were HIV positive, of whom 54% were diverted to the health system. 38% said they had never been tested for HIV. Factors associated with having a previous HIV test were: being over 20 years old; being HSH, SW, or PWID; being foreign; having used condoms at the last penetration; having had an STI in the last year; and having been in jail. CONCLUSIONS: The results show the great potential of these services to reach people who have never been tested for HIV. The HIV cases are concentrated in vulnerable groups. It is necessary to improve the linkage of diagnosed people to health care services.

OBJETIVO: La red COBATEST (red europea de centros comunitarios de cribado del VIH) permite la recogida estandarizada de información para monitorizar las pruebas de VIH realizadas en estos centros. El objetivo fue describir el rendimiento de los centros españoles de la red COBATEST y el perfil epidemiológico de sus usuarios, y analizar los factores asociados a tener una prueba de VIH anterior. METODOS: Análisis descriptivo de los datos recogidos en los 19 centros españoles que usaron las herramientas comunes de recogida de información durante el año 2015. Se realizó un modelo multivariado de regresión logística para identificar factores asociados a tener una prueba anterior. RESULTADOS: Se realizaron un total de 6.077 pruebas rápidas de VIH y de éstas un 1,8% fueron reactivas [3,0% HSH (hombres que tienen sexo con hombres); 3,1% TS (trabajadores/as sexuales); 2,3% UDI (usuarios/as de drogas inyectables); 0,6% heterosexuales)]. El 82,1% de los reactivos se realizaron la prueba confirmatoria y 94 (97,9%) fueron diagnosticados de VIH siendo derivados al sistema sanitario el 54%. El 38% afirmó no haberse realizado nunca la prueba del VIH. Los factores asociados a tener una prueba del VIH anterior son: ser mayor de 20 años; ser HSH, TS, o UDI; ser extranjero; haber usado preservativo en la última penetración; haber tenido una ITS en el último año; y haber estado en prisión. CONCLUSIONES: Los resultados muestran el gran potencial de estos servicios para llegar a personas que nunca se han realizado la prueba del VIH. Los casos de VIH se concentran en colectivos vulnerables. Es necesario mejorar la vinculación de las personas diagnosticadas a los servicios sanitarios.

Heterogeneity of community-based voluntary, counselling and testing services for HIV in Europe: the HIV-COBATEST survey.

This study reports the first EU-wide survey of community-based voluntary counselling and testing services (CBVCTs), with the following aims: to assess the current availability of CBVCTs and how the concept of CBVCT is understood; to describe CBVCT modalities and strategies; and to measure the use of rapid tests within CBVCTs. A cross-sectional survey was conducted in two different key informant groups: HIV/AIDS National Focal Points (NFPs) and CBVCTs from the EU and European Free Trade Association (EFTA) countries. A definition of CBVCTs was reached. Descriptive and correspondence analyses were performed. The COBATEST survey was answered by NFPs from 25 of the 32 EU/EFTA countries (response rate of 78.1%), and by 55 CBVCTs in 22 different countries. CBVCTs were functioning in nearly all the responding countries, but only 56% explicitly included them in their national strategic plans. In those that had CBVCTs, the NFPs often lacked reliable information on CBVCTs. In general, CBVCTs in Europe are focused mainly on men who have sex with men, are primarily peer-driven and highly communitarised, whereas CBVCTs targeting other at-risk populations are more medicalised. In addition, the oral test is under-used and in general test performance is highly medicalised. Results show that there is a wide heterogeneity of CBVCTs in Europe, which is varied and highly medicalised. The data provided in this study in conjunction with other deliverables produced by the COBATEST project should contribute to the development of standardised indicators to enable comparisons over time and eventually improve the effectiveness of CBVCTs across Europe.

Level of impact on the public health of universal human immunodeficiency virus screening in an Emergency Department. / Estudio del impacto a nivel de salud pública del cribado universal del virus de la inmunodeficiencia humana en un servicio de Urgencias.

INTRODUCTION: The aim of this study was to determine the prevalence of HIV and the acceptability of rapid testing in an emergency department (ED), Barcelona (6/07/2011 to 8/03/2013). METHOD: A convenience sample was used, depending on nurse availability in the ED. Participants signed an informed consent. Results were confirmed by conventional methods. RESULT: A total of 2,140 individuals were offered testing, and 5% rejected taking part (107/2,140). Three subjects (3/2,033 [0.15%]) had confirmed reactive test. Individuals with a higher education were more likely to perform a rapid HIV test in ED (P<.005). CONCLUSION: A low prevalence of new HIV diagnoses was found among participants, although there was a high acceptability rate to perform rapid testing in the ED.

Economic evaluation of HIV testing for men who have sex with men in community-based organizations - results from six European cities.

The non-decreasing incidence of HIV among men who have sex with men (MSM) has motivated the emergence of Community Based Voluntary Counselling and Testing (CBVCT) services specifically addressed to MSM. The CBVCT services are characterized by facilitated access and linkage to care, a staff largely constituted by voluntary peers, and private not-for-profit structures outside the formal health system institutions. Encouraging results have been measured about their effectiveness, but these favourable results may have been obtained at high costs, questioning the opportunity to expand the experience. We performed an economic evaluation of HIV testing for MSM at CBVCT services, and compared them across six European cities. We collected retrospective data for six CBVCT services from six cities (Copenhagen, Paris, Lyon, Athens, Lisbon, and Ljubljana), for the year 2014, on the number of HIV tests and HIV reactive tests, and on all expenditures to perform the testing activities. The total costs of CBVCTs varied from 54,390€ per year (Ljubljana) to 245,803€ per year (Athens). The cost per HIV test varied from to 41€ (Athens) to 113€ (Ljubljana). The cost per HIV reactive test varied from 1966€ (Athens) to 9065€ (Ljubljana). Our results show that the benefits of CBVCT services are obtained at an acceptable cost, in comparison with the literature (values, mostly from the USA, range from 1600$ to 16,985$ per HIV reactive test in clinical and non-clinical settings). This result was transversal to several European cities, highlighting that there is a common CBVCT model, the cost of which is comparable regardless of the epidemiological context and prices. The CBVCT services represent an effective and "worth it" experience, to be continued and expanded in future public health strategies towards HIV.

COBA-Cohort: a prospective cohort of HIV-negative men who have sex with men, attending community-based HIV testing services in five European countries (a study protocol).

INTRODUCTION: Community-based voluntary counselling and testing (CBVCT) services for men who have sex with men (MSM) can reach those most-at-risk and provide an environment for gay men that is likely to be non-stigmatising. Longitudinal data on the behaviour of HIV-negative MSM are scarce in Europe. The aim of this protocol, developed during the Euro HIV Early Diagnosis And Treatment (EDAT) project, is to implement a multicentre community-based cohort of HIV-negative MSM attending 15 CBVCT services in 5 European countries. RESEARCH OBJECTIVES: (1) To describe the patterns of CBVCT use, (2) to estimate HIV incidence, and to identify determinants of (3) HIV seroconversion and (4) HIV and/or sexually transmitted infection (STI) test-seeking behaviour. METHODS AND ANALYSIS: All MSM aged 18 years or over and who had a negative HIV test result are invited to participate in the COmmunity-BAsed Cohort (COBA-Cohort). Study enrolment started in February 2015, and is due to continue for at least 12 months at each study site. Follow-up frequency depends on the testing recommendations in each country (at least 1 test per year). Sociodemographic data are collected at baseline; baseline and follow-up questionnaires both gather data on attitudes and perceptions, discrimination, HIV/STI testing history, sexual behaviour, condom use, and pre- and post-exposure prophylaxis. Descriptive, exploratory and multivariate analyses will be performed to address the main research objectives of this study, using appropriate statistical tests and models. These analyses will be performed on the whole cohort data and stratified by study site or country. ETHICS AND DISSEMINATION: The study was approved by the Public Health authorities of each country where the study is being implemented. Findings from the COBA-Cohort study will be summarised in a report to the European Commission, and in leaflets to be distributed to study participants. Articles and conference abstracts will be submitted to peer-reviewed journals and conferences.

Implementation of rapid HIV and HCV testing within harm reduction programmes for people who inject drugs: a pilot study.

UNLABELLED: Including HCV and HIV rapid tests in harm reduction programmes (HRP) for people who inject drugs (PWID) can increase detection of these infections in high-risk populations who do not seek conventional health care. AIMS: To assess acceptability and feasibility of rapid HIV and HCV tests in HRP; to identify HIV and HCV prevalence rates in HRP; to identify the percentage of PWID with a reactive test that attend hospital for confirmation and follow-up. Rapid oral tests for HCV and HIV were offered to users of 13 HRP from both mobile units and facility-based centres. A total of 172 HCV and 198 HIV tests were performed, with a refusal rate of 1.7% and 10.4%, respectively. Injectors made up 64.9% of all drug users and 35.1% did not inject drugs. Overall, 20.3% of HCV tests and 2.5% of HIV test were reactive. Only 24 of the 35 reactive HCV could be confirmed (68.6%) and one was false-negative. Of the five HIV reactive cases, only two could be confirmed (40%) with 1 false-positive case. Acceptability of rapid HIV and HCV tests among HRP users was high. The usefulness of oral rapid tests in HRP has been demonstrated, especially in mobile HRP.

Approval and potential use of over-the-counter HIV self-tests: the opinion of participants in a street based HIV rapid testing program in Spain.

HIV self-testing, not yet available in Spain, is a strategy thought to be able to increase the number of people tested and testing frequency. 3,373 attenders of a street-based HIV rapid-testing program gave their opinion on authorizing over-the-counter self-tests and a potentially shorter lead time if self-tests were available. 88.0 % of participants were in favor of authorization, 9.2 % had no clear opinion and 2.8 % were against. 54.6 % of men who have sex with men (MSM), 38.4 % of men who have sex with women and 36.3 % of women acknowledged a potential for lead time reduction. Potential lead time was associated with being ≥35 years, having a university degree, never injected drugs, previous HIV tests and being concerned about an HIV positive result, and in MSM, also having ≥5 partners. Self-testing seems a promising strategy for Spain: its authorization is supported by nearly all potential users and almost three quarters of MSM would have opted to advance their current testing if self-tests were available.