María Luisa Mateos

Hepatitis E Virus: Relevance in Blood Donors and Risk Groups

Background and Objectives: Hepatitis E virus (HEV) infection usually causes acute self–limited disease. HEV is associated with faecal–contaminated drinking water, but other vectors, such as blood, are possible. The aim of this study was to investigate the prevalence of HEV in blood donors and in two groups at high risk for parenteral infections, namely, haemodialysis patients, and children infected with HCV via blood transfusion. Materials and Methods: We investigated the prevalence of anti–HEV in 863 blood donors, 63 haemodialysis patients, and 42 children infected post transfusion with HCV. Results: The prevalence rates were 2.8, 6.3%, and zero, respectively. Conclusions: (1) The incidence of HEV in Spain is similar to that in other Western European countries, and (2) HEV is probably not transmitted parenterally to children.

Pegylated interferon α2b plus ribavirin for the treatment of chronic hepatitis C in HIV-infected patients

Background: Hepatitis C virus (HCV) and HIV coinfection constitutes an important epidemiological and clinical problem. We evaluated the safety and efficacy of Pegylated interferon α2b (Peg-IFN) and a fixed dose of ribavirin in the treatment of chronic hepatitis C in HIV coinfection. Methods: Open, prospective study in HCV–HIV coinfected patients with persistently elevated alanine aminotransferase (ALT) levels and a liver biopsy showing either portal or bridging fibrosis. Therapy included Peg-IFN (50 μg weekly) with ribavirin 800 mg for 48 weeks. The primary end point was sustained virological response (SVR). Univariate and multivariate analyses were performed to determine factors associated with response. Results: By intent-to-treat analysis, 11 of 35 patients (31%) reached SVR. SVR was significantly better for genotypes 2/3 than for genotype 1 (54% versus 21%; P < 0.05). By multivariate logistic regression analysis, only a non-1 genotype was an independent factor for SVR [odds ratio (OR), 6; 95% confidence interval (CI), 1.1–31.7; P < 0.005]. A decrease of at least 1.5 log10 HCV RNA at week 12 of therapy was highly predictive of SVR (OR, 49.9; 95% CI, 4.9–508.2; P < 0.001). Most patients developed adverse events, although only six patients (17%) discontinued treatment due to toxicity. Conclusions: The combination of low doses of Peg-IFN plus a fixed dose of ribavirin resulted in a rate of SVR similar to that obtained with higher doses of the drugs in HIV-infected patients and lower than those obtained in non-HIV patients. Response at week 12 may be useful to help guide therapy in HCV–HIV co-infected patients.

Safety and anti-HCV effect of prolonged intravenous silibinin in HCV genotype 1 subjects in the immediate liver transplant period

BACKGROUND & AIMS Reinfection of the graft is the rule in patients with HCV cirrhosis undergoing liver transplantation, and HCV-RNA reaches pre-transplantation levels within the first month. Short-term intravenous silibinin monotherapy is safe and shows a potent in vivo anti-HCV effect. We aimed at evaluating the safety and antiviral effect of prolonged intravenous silibinin, started immediately before liver transplantation. METHODS Single centre, prospective, pilot study, to assess the safety and effect on HCV-RNA kinetics during at least 21 days of intravenous silibinin monotherapy (20 mg/kg/day) in 9 consecutive HCV genotype 1 subjects, in comparison to a control, non-treated group of 7 consecutive prior transplanted subjects under the same immunosuppressive regimen (basiliximab, steroids, delayed tacrolimus, micophenolate). RESULTS Intravenous silibinin led to significant, maintained and progressive HCV-RNA decreases (mean HCV-RNA drop at week 3, -4.1 ± 1.3 log(10)IU/ml), and lack of viral breakthrough during administration. Four patients (44%) reached negative HCV-RNA, maintained during silibinin treatment, vs. none in the control group, but HCV-RNA relapsed in all of them after a median of 21 days (16-28), following silibinin withdrawal. Partial responders to silibinin showed marked decreases in HCV-RNA when compared to controls, but lower than complete responders. There were no clinical adverse effects, and silibinin led to asymptomatic transient hyperbilirubinemia (week 2, 4.2 ± 2.2 vs. 2.5 ± 3.6 mg/dl; p=0.02). CONCLUSIONS Prolonged intravenous silibinin monotherapy was safe in the immediate liver transplantation period, leading to a potent and time dependent antiviral effect and lack of HCV-RNA breakthrough during administration. However, HCV-RNA rebounded after withdrawal, and silibinin monotherapy did not avoid reinfection of the graft.

Hepatitis aguda E en Madrid: descripción de 18 casos

Resumen Introduccion En paises subdesarrollados, el virus de la hepatitis E (VHE) es el principal causante de las hepatitis no-A no-B no-C epidemicas de transmision enterica. Sin embargo, en estos ultimos anos se han descrito casos de hepatitis esporadicas en Europa, sin que se encuentre como antecedente epidemiologico el viaje a paises endemicos. Objetivo y pacientes Presentamos 12 casos de hepatitis E esporadicas autoctonas (no se encontro ningun factor de riesgo epidemiologico) y 6 importadas (antecedente de viaje a Nepal, Santo Domingo, China, Brasil y Ecuador) estudiados entre junio de 1999 y diciembre de 2005. Resultados El diagnostico de hepatitis E se realizo en pacientes con sintomas de hepatitis aguda que presentaban anticuerpos IgG e IgM anti-VHE en sangre. El VHE es la causa de algunas hepatitis agudas en paises desarrollados que antes se etiquetaban como de etiologia desconocida. Conclusiones La hepatitis E debe incluirse en el diagnostico diferencial de hepatitis aguda incluso en pacientes sin antecedentes de viajes a zonas endemicas.

Dried Blood as an Alternative to Plasma or Serum for Trypanosoma cruzi IgG Detection in Screening Programs

ABSTRACT Trypanosoma cruzi serological screening is recommended for people potentially exposed to this parasite in countries where Trypanosoma cruzi is endemic and those where it is not endemic. Blood samples on filter paper may be a practical alternative to plasma/serum for antibody detection. Using the Architect Chagas assay, we detected the presence of IgG against T. cruzi in matched serum and dried blood spots (DBS) collected from 147 patients residing in Madrid, Spain, who had potential previous exposure to T. cruzi. The κ statistic for the DBS/serum proportion of agreement for the detection of antibodies against T. cruzi was 0.803, considering an S/CO (assay result unit; chemiluminescent signal from the sample [S] divided by the mean chemiluminescent signal for the three calibrators used in the test [CO]) cutoff value of ≥1.00. The relative sensitivity of the Architect test using DBS increased from 95.2% to 98.8% when the cutoff was lowered from ≥1.00 to ≥0.88, while the relative specificity decreased from 84.1% to 71.6%. Overall, the median S/CO values for DBS were significantly lower than those for serum (2.6 versus 6.5; P < 0.001). Discrepancies that occurred with the use of DBS included 10 false positives (with low S/CO values in 9 cases [median, 2.13]) and 4 false negatives, with mean S/CO values of 0.905 (gray zone). Using DBS plus a highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) may be a simple and reliable method for detecting IgG against T. cruzi when blood sampling by venipuncture is not feasible. This method may also reduce the false-negative rates observed with some rapid diagnostic tests. The lower relative sensitivity compared to the reference method may be increased by lowering the optical density threshold.

Detección y genotipado del virus del papiloma humano de alto riesgo en muestras de lesiones cervicales

INTRODUCTION This study investigates the relationship between various human papillomavirus (HPV) genotypes and the results of cytological and histological analysis of cervical samples using two complementary assays for HPV detection (hybrid capture and PCR). We studied the impact of HPV genotype on the presence of pre-cancerous cervical lesions and cervical cancer, as well as the association between HPV viral load and the presence of high-risk HPV as determined by PCR. METHODS A total of 272 women were studied. Most of them presented cellular alterations consistent with cervical lesions due to HPV and all had high-risk HPV as detected by hybrid capture testing. Histological studies were undertaken, and HPV genotyping by PCR based on microarrays was performed. RESULTS HPV-DNA was not detected or genotypes could not be identified by PCR in 22.06% of the patients. Genotype 16 and/or 18 was detected in 33% of 212 patients. Mixed infections with several genotypes were found in 25% of patients. The histological lesions associated with the various genotypes were as follows: genotype 16 and/or 18. were detected in 55.73% of the 61 patients with H-SIL and cancer, whereas these genotypes were detected in only 7.9% and 22% of women with ASCUS and L-SIL (P < 0.05). Viral load was less than 3 pg/mL in 12.13% of the women studied. In this group of patients, high-risk HPV was present in 39.39%. In the group of patients who had a viral load greater than 3 pg/mL, high risk-HPV was detected in 77.4% (P < 0.05). CONCLUSIONS Genotypes 16 and/or 18 were detected in most patients with a diagnosis of H-SIL. Other high-risk-HPV genotypes were much less prevalent. Hybrid capture testing is a useful screening test. PCR was effective for identifying genotypes 16 and 18. Histological and cytological findings in cervical samples should be interpreted together with high-risk HPV detection.

Relevance of cutoff on a 4th generation ELISA performance in the false positive rate during HIV diagnostic in a low HIV prevalence setting

BACKGROUND Despite the high specificity of fourth-generation enzyme immunoassays (4th-gen-EIA) for screening during HIV diagnosis, their positive predictive value is low in populations with low HIV prevalence. Thus, screening should be optimized to reduce false positive results. OBJECTIVES The influence of sample cutoff (S/CO) values by a 4th-gen-EIA with the false positive rate during the routine HIV diagnosis in a low HIV prevalence population was evaluated. STUDY DESIGN A total of 30,201 sera were tested for HIV diagnosis using Abbott Architect® HIV-Ag/Ab-Combo 4th-gen-EIA at a hospital in Spain during 17 months. Architect S/CO values were recorded, comparing the HIV-1 positive results following Architect interpretation (S/CO≥1) with the final HIV-1 diagnosis by confirmatory tests (line immunoassay, LIA and/or nucleic acid test, NAT). ROC curve was also performed. RESULTS Among the 30,201 HIV performed tests, 256 (0.85%) were positive according to Architect interpretation (S/CO≥1) but only 229 (0.76%) were definitively HIV-1 positive after LIA and/or NAT. Thus, 27 (10.5%) of 256 samples with S/CO≥1 by Architect were false positive diagnose. The false positive rate decreased when the S/CO ratio increased. All 19 samples with S/CO ≤10 were false positives and all 220 with S/CO>50 true HIV-positives. The optimal S/CO cutoff value provided by ROC curves was 32.7. No false negative results were found. CONCLUSIONS We show that very low S/CO values during HIV-1 screening using Architect can result HIV negative after confirmation by LIA and NAT. The false positive rate is reduced when S/CO increases.

Recent contributions for improving sensitivity in chiral CE

The flexibility and versatility of the chiral CE are unrivaled and the same instrumentation can be used to separate a diverse range of analytes, both large and small molecules, whether charged or uncharged. However, one of the disadvantages is generally thought to be the poor sensitivity of ultraviolet (UV) detection, which is the most popular among CE detectors. This review focuses on methodologies and applications regarding improvements of sensitivity in chiral CE published in the last 2 years (June 2015 until May 2017). This contribution continues to update this series of biannual reviews, first published in Electrophoresis in 2006. The main body of the review brings a survey of publications organized according to different approaches to detect a low amount of analytes, either by sample treatment procedures or by in‐capillary sample preconcentration techniques, both using UV detection, or even by employing detection systems more sensitive than UV absorption, such as LIF or MS. This review provides comprehensive tables listing the new approaches in sensitive chiral CE with categorizing by the fundamental mechanism to enhance the sensitivity, which provide relevant information on the strategies employed. The concluding remarks in the final part of the review evaluate present state of art and the trends for sensitivity enhancement in chiral CE.

Evaluation of Cobas Ampliprep Automated Nucleic Acid Extraction for Genotyping Human Papillomavirus (HPV) by the Roche linear array HPV Test

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