Evaluation of the Xpert HCV VL Fingerstick point-of-care assay and dried blood spot HCV-RNA testing as simplified diagnostic strategies among people who inject drugs in Catalonia, Spain.

BACKGROUND: Catalonia requires decentralized and simplified strategies for the diagnosis of viremic HCV infection among people who inject drugs (PWID). We aimed to perform a direct comparison of the diagnostic performance between two, single-step strategies for the screening and diagnosis of viremic HCV infection in PWID attending a drug consumption room (DCR) in Barcelona: i) on-site HCV-RNA testing using the point-of-care test (PoCT) Xpert HCV VL Fingerstick; and ii) on-site dried blood spots (DBS) collection for HCV-RNA testing at the laboratory (in-house assay). Additionally, we aimed to assess participants' preferences in receiving HCV-RNA testing results and feasibility of same-day delivery of PoCT results. METHODS: The real-world, clinical performance of these two strategies was established in comparison with the reference method (HCV viral load testing with the Xpert HCV Viral Load assay at the laboratory from venous plasma collected at the DCR). HCV genotypes/subtypes and HIV status were also determined by sequencing and serology, respectively. A questionnaire including preferences regarding the delivery of test results was administered. RESULTS: The prevalence of HCV-RNA was 63.0% (of which 25.8% were co-infected with HIV). The RNA-PoCT showed a sensitivity of 98.4% for detectable viral loads (>4 IU/mL) and of 100% for quantifiable viral loads (≥10, ≥1000 and ≥3000 IU/mL). For the DBS-based assay, a sensitivity of 93.7% was obtained for detectable viral loads, 96.7% for the quantifiable ≥10 IU/mL threshold, and 98.3% for both the quantifiable ≥1000 and ≥3000 IU/mL thresholds. No significant differences were detected between the sensitivity values of these two strategies, and the specificity was 100% in both cases. Half of the participants preferred to receive the HCV-RNA result on the same day, and 80% of participants received their RNA-PoCT results on the same day. CONCLUSIONS: Both the HCV-RNA PoCT and the DBS-based assay are highly reliable tools for the simplified diagnosis of viremic HCV infection among current PWID. These strategies allow for on-site sample collection and delivery of test results, facilitating decentralized care in harm reduction services.

Reliable resolution of ambiguous hepatitis C virus genotype 1 results with the Abbott HCV Genotype Plus RUO assay.

Accurate subtyping of hepatitis C virus genotype 1 (HCV-1) remains clinically and epidemiologically relevant. The Abbott HCV Genotype Plus RUO (GT Plus) assay, targeting the core region, was evaluated as a reflex test to resolve ambiguous HCV-1 results in a challenging sample collection. 198 HCV-1 specimens were analysed with GT Plus (38 specimens with and 160 without subtype assigned by the Abbott RealTime Genotype II (GT II) assay targeting the 5'NC and NS5B regions). Sanger sequencing of the core and/or NS5B regions were performed in 127 specimens without subtype assignment by GT II, with "not detected" results by GT Plus, or with mixed genotypes/subtypes. The remaining GT Plus results were compared to LiPA 2.0 (n = 45) or just to GT II results if concordant (n = 26). GT Plus successfully assigned the subtype in 142/160 (88.8%) samples. "Not detected" results indicated other HCV-1 subtypes/genotypes or mismatches in the core region in subtype 1b. The subtyping concordance between GT Plus and either sequencing or LiPA was 98.6% (140/142). Therefore, combined use of GT II and GT Plus assays represents a reliable and simple approach which considerably reduced the number of ambiguous HCV-1 results and enabled a successful subtyping of 98.9% of all HCV-1 samples.

Community-based screening of hepatitis C with a one-step RNA detection algorithm from dried-blood spots: Analysis of key populations in Barcelona, Spain.

Alternative strategies are required to enhance the diagnosis of silent hepatitis C virus (HCV) infections in key populations at risk. Among them, HCV prevalence and bio-behavioural data are scarce for HIV-negative men who have sex with men (MSM) and men and trans-women sex workers. We sought to describe and assess the potential benefits of a community-based one-step HCV screening and confirmatory strategy for these populations in Barcelona. The screening strategy based on a real-time RT-PCR assay for HCV-RNA detection in dried-blood spots (DBS) was validated and implemented in addition to an antibody point-of-care test in a community centre. HCV prevalence was assessed, and bio-behavioural data were collected. The molecular assay was precise, reproducible, sensitive and specific. Four HIV-negative MSM reported being currently infected (0.75% HCV self-reported prevalence). Implementation of DBS testing was easy, and acceptability was >95%, but no silent HCV case was diagnosed (N = 580). High-risk sexual practices and drug use for sex were reported frequently. HIV prevalence was 4.7% in MSM and 10% in sex workers. Self-reported prevalence of other STIs ranged from 11.3% to 36.2%. In conclusion, HCV-RNA testing in DBS showed a good performance, but the assessed one-step strategy does not seem beneficial in this setting. Although no silent HCV infections were detected, the observed high-risk behaviours and prevalence of other STIs suggest that HCV spread should be periodically monitored among these populations in Barcelona by means of behavioural surveillance, rapid antibody testing and molecular confirmation in DBS.

Prevalence and distribution of hepatitis C virus genotypes in Spain during the 2000-2015 period (the GEHEP 005 study).

We report the largest study on the prevalence and distribution of HCV genotypes in Spain (2000-2015), and we relate them with clinical, epidemiological and virological factors. Patients from 29 hospitals in 10 autonomous communities (Andalusia, Aragon, Castilla-Leon, Catalonia, Galicia, Canary Islands, Madrid Community, Valencian Community, Murcia Region and Basque Country) have been studied. Annual distribution of HCV genotypes and subtypes, as well as gender, age, transmission route, HIV and/or HBV coinfection, and treatment details were recorded. We included 48595 chronically HCV-infected patients with the following characteristics: median age 51 years (IQR, 44-58), 67.9% male, 19.1% HIV-coinfected, 23.5% HBV-coinfected. Parenteral transmission route was the most frequent (58.7%). Genotype distribution was 66.9% GT1 (24.9% subtype 1a and 37.9% subtype 1b), 2.8% GT2, 17.3% GT3, 11.4% GT4 and 0.1% GT5 and 0.02% GT6. LiPA was the most widely HCV genotyping test used (52.4%). HCV subtype 1a and genotypes 3 and 4 were closely associated with male gender, parenteral route of infection and HIV and HBV coinfection; in contrast, subtype 1b and genotype 2 were associated with female gender, nonparenteral route and mono-infection. Age was related to genotype distribution, and different patterns of distribution and biodiversity index were observed between different geographical areas. Finally, we describe how treatment and changes in transmission routes may have affected HCV genotype prevalence and distribution patterns. We present the most recent data on molecular epidemiology of hepatitis C virus in Spain. This study confirms that genotype distributions vary with age, sex, HIV and HBV coinfection and within geographical areas and epidemiological groups.

Forma de presentación atípica de panencefalitis esclerosante subaguda del sarampión en el adulto / Atypical presentation of adult-age onset subacute sclerosing panencephalitis

No disponible

Epidemiological features and determinants for Chlamydia trachomatis infection among women in Catalonia, Spain.

We determined the prevalence of Chlamydia trachomatis (CT) infection and risk factors for its acquisition by women aged less than 35 years who were attending sexual health centres in the province of Barcelona in 2007. A convenience sample of 397 adolescents and young adult women aged between 16 and 35 years, stratified by age and recruitment site, were tested using realtime polymerase chain reaction by CT DNA. A standardized questionnaire was used to collect epidemiological and behavioural data on the participants, which were analysed in a multivariate logistic regression model. The overall prevalence of CT was 4%, significantly higher in those under 25 years of age (5.8% versus 1.6% over 25). The independent risk factors for CT infection were: foreign origin (odds ratio [OR] 4.7; confidence interval [CI] 1.02-21.8), having had a sexual partner in the last three months (OR 4.59; CI 1.16-18.08) and tobacco use the last 12 months (OR 6.38; CI 1.16-34.93). In conclusion, this is the first study performed in Catalonia showing a high prevalence of CT in young women, consistent with trends in the rest of Europe. Systematic monitoring of CT infection in sentinel populations such as this will inform future targeted screening programmes in our setting.

Recently acquired HIV infection in Spain (2003-2005): introduction of the serological testing algorithm for recent HIV seroconversion.

BACKGROUND: Identification of recent HIV infections provides a description of the current pattern of HIV transmission and, consequently, can help to design better preventive interventions. Our study shows the first implementation in Spain of the Serologic Testing Algorithm for Recent HIV Seroconversion (STARHS) strategy. We assess the viability of introducing STARHS in our setting and describe the frequency and epidemiological characteristics of recent infections (RIs). METHODS: Between 2003 and 2005, HIV-positive blood samples drawn for diagnostic purposes were collected from 28 Spanish laboratories to be tested using STARHS. Samples from patients with a previous HIV diagnosis, age <18 years, <200 CD4 cells/microL or clinical AIDS criteria were excluded from the analysis. RESULTS: A total of 660 (19.2%) samples were classified as RI. Most people identified with RI were male (79.8%) with a median age of 33.1 years, and 62.5% occurred among men who have sex with men (MSM). Immigrants made up 26.5% of individuals identified with RIs, with 48.7% coming from South America. Among the individuals with RI, at least 16.5% had reported another sexually transmitted infection (STI) during the year before the HIV diagnosis. CONCLUSION: The study shows that the implementation of STARHS in our setting is feasible and has highlighted important features of the local HIV epidemic, such as the ongoing spread of HIV among MSM, the potential role of STIs in RIs and the vulnerability of immigrants as a new target population.

Detection of hepatitis C virus antibodies in oral fluid specimens for prevalence studies.

Within the framework of hepatitis C virus (HCV) prevalence monitoring, we evaluated oral fluid (OF), which is richer in IgG than whole saliva, as a possible alternative to serum for the detection of HCV antibodies. Paired OF and serum samples were collected from 90 individuals, including 45 HCV-positives and 45 HCV-negatives. The detection of HCV antibodies in both serum and OF was performed using the Ortho HCV 3.0 SAVe enzyme-linked immunosorbent assay (ELISA) (Ortho-Clinical Diagnostics, Inc., Raritan, NJ), but a modified, more sensitive protocol was used to process OF. The sensitivity and specificity of this assay were 86.67% (95% confidence interval (CI): 72.51-94.46%) and 100% (95% CI: 90.20-99.80%) in OF and 100% in serum. The correlation obtained between both types of clinical specimens was excellent (k: 0.87, 95% CI: 0.66-1.07). However, the negative predictive value (NPV) of the assay in OF decreased with the prevalence of HCV infection in the population studied. Our results suggest that the modified Ortho HCV 3.0 SAVe ELISA is suitable for the detection of HCV antibodies in OF for epidemiological studies. Using this assay, we observed an unadjusted anti-HCV prevalence of 78.6% among a population of intravenous drug users; when adjusted to account for assay sensitivity, this prevalence may be closer to 90%.

Mycobactericidal and tuberculocidal activity of Korsolex AF, an amine detergent/disinfectant product.

The mycobactericidal and tuberculocidal activities of Korsolex AF against Mycobacterium tuberculosis, Mycobacterium avium-Mycobacterium intracellulare (MAI), Mycobacterium kansasii and Mycobacterium chelonae were determined using quantitative suspension and carrier tests. The effects of organic load and hard water were also considered. A clinical isolate of MAI was the most resistant of the four test organisms. A 2% solution had good mycobactericidal and tuberculocidal activities after 30 min of exposure. Although further evaluation using European standard tests is necessary, we conclude that Korsolex AF appears to be a promising product for the disinfection of hospital instruments contaminated with mycobacteria.

Comparison of human herpesvirus 8 and Epstein-Barr virus seropositivity among children in areas endemic and non-endemic for Kaposi's sarcoma.

Human herpesvirus 8 (HHV-8) is the etiologic agent of Kaposi's sarcoma (KS). Several studies indicate horizontal HHV-8 transmission among children in areas where KS is endemic, but few studies have assessed acquisition of HHV-8 by children in low seroprevalence areas. Antibody screening was carried out for HHV-8 and Epstein-Barr virus (EBV) on 787 serum specimens from children living in two areas where HHV-8 is not endemic, the United States (US) and Germany, and on 184 specimens from children living in a KS-endemic area (Nigeria). For children in the US and Germany, the results showed low HHV-8 seroprevalence rates (3-4%). However, US children aged 6 months to 5 years had higher HHV-8 antibody titers than did 6-17-year-old children (P < 0.01), a finding consistent with more recent infections being detected in the younger children. Compared with seroprevalence rates and antibody titers in US and German children, those in Nigerian children were significantly higher, and seroprevalence increased with age. There was no evidence of cross-reactivity between assays for HHV-8 and EBV, despite the genetic similarity of these two herpesviruses. The data indicate that HHV-8 transmission among children where HHV-8 is not endemic occurs, but is uncommon. The findings also suggest that HHV-8 antibodies, as measured by current tests, may not persist for long periods in populations at low risk for KS and that vertical transmission is rare, although longitudinal studies are necessary to address directly these issues.

Assessment of Acinetobacter baumannii susceptibility to antiseptics and disinfectants.

Disinfection and antisepsis are of primary importance in controlling outbreaks of Acinetobacter baumannii, a nosocomial pathogen that frequently shows multiple antibiotic resistance. In this study we assessed the susceptibility of nine A. baumannii strains isolated during a sustained intensive care unit outbreak, to several antiseptics and disinfectants based on European Standards. While the tested strains showed diverse antibiotic resistance patterns, they were equally sensitive to the biocides assessed in vitro. We observed neither evidence of development of resistance to biocides over time, nor a correlation between resistance to antibiotics and a decreased susceptibility to antiseptics or disinfectants.

In-use evaluation of Perasafe compared with Cidex in fibreoptic bronchoscope disinfection.

The mycobactericidal activity of Perasafe (0.26% peracetic acid) was compared with that of Cidex (2% alkaline glutaraldehyde) by an in-use test. Fibreoptic bronchoscopes were artificially contaminated with Mycobacterium tuberculosis or Mycobacterium avium-intracellulare in sputum and, after manual pre-cleaning with a neutral soap, 10 and 20 min disinfection periods were tested. Perasafe was as effective as Cidex, thus requiring a 10 min disinfection period against M. tuberculosis and 20 min against M. avium-intracellulare. The results demonstrate that Perasafe is an effective disinfectant for use in reprocessing fibreoptic bronchoscopes.

In-vitro evaluation of Perasafe compared with 2% alkaline glutaraldehyde against Mycobacterium spp.

Quantitative suspension and carrier tests were used to compare the activity of Perasafe and Cidex against Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, Mycobacterium fortuitum, and Mycobacterium chelonae. The interference of an organic load, and of hard water was also considered. Both agents achieved reductions exceeding 10(5)-fold within 20 and 30 min for all the strains tested. Perasafe is thus mycobactericidal and a viable alternative to Cidex for intermediate or high-level disinfection.

Assessment of in-vitro efficacy of 1% Virkon against bacteria, fungi, viruses and spores by means of AFNOR guidelines.

Peroxygenic acid, under the brand name Virkon, has unleashed great debate following contradictory reports of its efficacy and spectrum of activity. The aim of this study was to test the biocidal activity of the compound against 10 different micro-organisms, following standard in-vitro test procedures. Bactericidal, fungicidal and sporicidal activities were determined using quantitative suspension and germ carrier tests and virucidal activity was assessed using a simple dilution suspension test, following the Association Française de Normalisation (AFNOR) guidelines. One percent Virkon demonstrated bactericidal activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Enterococcus hirae and Mycobacterium smegmatis in the suspension test and against P. aeruginosa, E. coli, S. aureus and E. hirae in the carrier test. One percent Virkon showed virucidal activity against poliovirus in the suspension test. However, this concentration did not comply with sporicidal and fungicidal activity guidelines. In conclusion, 1% Virkon is effective only against vegetative bacteria, yeasts and viruses, and should therefore be considered a low-level disinfectant.