Silvia García-Bujalance

Molecular Surveillance of HIV-1 in Madrid, Spain: a Phylogeographic Analysis

ABSTRACT The molecular epidemiology of HIV-1 is constantly changing, mainly as a result of human migratory flows and the high adaptive ability of the virus. In recent years, Spain has become one of Europes main destinations for immigrants and one of the western European countries with the highest rates of HIV-positive patients. Using a phylogeographic approach, we have analyzed the relationship between HIV-1 variants detected in immigrant and native populations of the urban area of Madrid. Our project was based on two coincidental facts. First, resistance tests were extended to naïve and newly diagnosed patients, and second, the Spanish government legislated the provision of legal status to many immigrants. This allowed us to obtain a large data set (n = 2,792) from 11 Madrid hospitals of viral pol sequences from the two populations, and with this unique material, we explored the impact of immigration in the epidemiological trends of HIV-1 variants circulating in the largest Spanish city. The prevalence of infections by non-B HIV-1 variants in the studied cohort was 9%, rising to 25% among native Spanish patients. Multiple transmission events involving different lineages and subsubtypes were observed in all the subtypes and recombinant forms studied. Our results also revealed strong social clustering among the most recent immigrant groups, such as Russians and Romanians, but not in those groups who have lived in Madrid for many years. Additionally, we document for the first time the presence of CRF47_BF and CRF38_BF in Europe, and a new BG recombinant form found in Spaniards and Africans is tentatively proposed. These results suggest that the HIV-1 epidemic will evolve toward a more complex epidemiological landscape.

Comparison of Imported Plasmodium ovale curtisi and P. ovale wallikeri Infections among Patients in Spain, 2005–2011

Sequencing data from Plasmodium ovale genotypes co-circulating in multiple countries support the hypothesis that P. ovale curtisi and P. ovale wallikeri are 2 separate species. We conducted a multicenter, retrospective, comparative study in Spain of 21 patients who had imported P. ovale curtisi infections and 14 who had imported P. ovale wallikeri infections confirmed by PCR and gene sequencing during June 2005–December 2011. The only significant finding was more severe thrombocytopenia among patients with P. ovale wallikeri infection than among those with P. ovale curtisi infection (p = 0.031). However, we also found nonsignificant trends showing that patients with P. ovale wallikeri infection had shorter time from arrival in Spain to onset of symptoms, lower level of albumin, higher median maximum core temperature, and more markers of hemolysis than did those with P. ovale curtisi infection. Larger, prospective studies are needed to confirm these findings.

Initial experience with imported Zika virus infection in Spain.

INTRODUCTION A considerable increase of imported Zika virus (ZIKV) infection has been reported in Europe in the last year. This is the result of the large outbreak of the disease in the Americas, along with the increase in the numbers of travellers and immigrants arriving from ZIKV endemic areas. METHODS A descriptive study was conducted in the Tropical Medicine Unit of Hospital La Paz-Carlos III in Madrid on travellers returning from an endemic area for ZIKV from January to April 2016. Demographic, clinical and microbiological data were analyzed. RESULTS A total of 185 patients were screened for ZIKV (59.9% women, median age of 37.7±10.3 years). Main purpose of the travel was tourism to Colombia, Brazil, and México. Just under three-quarters (73%) were symptomatic, mostly with fever and headache. A total of 13 patients (7% of those screened) were diagnosed with ZIKV infections, of which four of them were pregnant. All of them were symptomatic patients, the majority immigrants, and mainly from Colombia. Diagnostic tests were based on positive neutralization antibodies (8 cases, 61.6%) and a positive RT-PCR in different organic fluids (7 cases, 53.8%) The four infected pregnant women underwent a neurosonography every 3 weeks, and no alterations were detected. RT-PCR in amniotic fluid was performed in three of them, with negative results. One of the children has already been born healthy. CONCLUSIONS Our cases series represents the largest cohort of imported ZIKV to Spain described until now. Clinicians must increase awareness about the progression of the ZIKV outbreak and the affected areas so that they can include Zika virus infection in their differential diagnosis for travellers from those areas.

Persistence and infectivity of Zika virus in semen after returning from endemic areas: Report of 5 cases

BACKGROUND There are limited data about the persistence and infectivity of Zika virus in semen of symptomatic travelers returning from endemic areas and even less data in asymptomatic cases. OBJECTIVE We investigated the persistence and infectivity of ZIKA virus in semen in five patients with Zika virus infection returning to Spain from endemic areas. STUDY DESIGN We evaluated the epidemiological, clinical and virological characteristic of the five patients. In semen we detected ZIKA virus by PCR, partial sequencing and cell culture. We also performed phylogenetic analysis. RESULTS We detected Zika virus RNA (Asian lineage) by PCR in semen samples from day 14th to day 96th since the day of illness onset. Semen viral culture was positive for Zika virus in two patients at days of illness 30 and 69 by virus propagation. Phylogenetic analysis strongly suggested male to female sexual transmission in a couple returning from Maldives. CONCLUSION This case series confirms that Zika virus RNA can be detected in semen up to three months after infection. Viral culture of semen samples shows prolonged infectivity that can lead to sexual transmission of Zika virus.

Comparison between Sequence Analysis and a Line Probe Assay for Testing Genotypic Resistance of Human Immunodeficiency Virus Type 1 to Antiretroviral Drugs

ABSTRACT The purpose of this study was to compare a line probe assay (LiPA) with sequence analysis for the detection of mutations conferring resistance to nucleoside and non-nucleoside inhibitors in human immunodeficiency reverse transcriptase and protease inhibitors. The limitations for interpreting LiPA make it unacceptable for routine clinical practice.

Detection and quantification of the K103N mutation in HIV reverse transcriptase by pyrosequencing.

Prolonged treatment of human immunodeficiency virus (HIV)-infected patients with nonnucleoside reverse transcriptase inhibitors (NNRTIs) might result in the selection of resistant mutants, the most frequent being the K103N mutation in reverse transcriptase. Resistance mutations are routinely detected by Sanger sequencing of the whole viral population, which does not detect sequence variants with frequencies below 20%. We have developed a pyrosequencing approach for the analysis of codon 103 of the HIV reverse transcriptase gene in the circulating viral population that detects variants below the limit of conventional sequencing. The method was tested with samples from 5 controls (not exposed to NNRTIs), 6 from patients exposed to NNRTIs and having a K103N mutant virus population detected by conventional sequencing, and 9 from patients previously exposed to NNRTIs that had a wild-type virus population by conventional sequencing. In 7 of 9, samples the mutation could not be detected by either the standard assay or pyrosequencing, while in 2 samples persistence of the mutation could be detected by pyrosequencing. The method might be of practical use in detecting minority variants of HIV in the clinical setting, in epidemiological studies with large numbers of samples, or as a complement to more complex approaches.

Is etravirine and two nucleosides an option for HIV with an isolated K103N mutation

We report long-term virologic response to etravirine and tenofovir/emtricitabine in four HIV-1-infected patients who had prior standard genotypic resistance testing showing an isolated K103N mutation (three acquired, one transmitted). In three patients tested, the K103N mutation was detected in cellular HIV-1 DNA whereas remaining suppressed on etravirine plus tenofovir/emtricitabine.

Imported Plasmodium falciparum malaria in HIV-infected patients: a report of two cases.

As HIV becomes a chronic infection, an increasing number of HIV-infected patients are travelling to malaria-endemic areas. Association of malaria with HIV/AIDS can be clinically severe. Severe falciparum malaria is a medical emergency that is associated with a high mortality, even when treated in an Intensive Care Unit. This article describes two cases of HIV-positive patients, who returned from malaria-endemic areas and presented a parasitaemia > 5% of erythrocytes and clinical signs of severe falciparum malaria, both with > 350 CD4 cell count/μl, absence of chemoprophylaxis and successful response. Factors like drug interactions and the possible implication of anti-malarial therapy bioavailability are all especially interesting in HIV-malaria co-infections.

Emergence of VIM-1-producing Salmonella enterica serovar Typhimurium in a paediatric patient.

The blaVIM gene encodes a metallo-blactamase enzyme with activity against most b-lactams, including carbapenems (Queenan & Bush, 2007). The gene is most often found as an integron-borne gene cassette with high mobility among plasmids, chromosomes and other integrons. It was described initially in Pseudomonas aeruginosa (Lauretti et al., 1999) but has disseminated to the Enterobacteriaceae (Queenan & Bush, 2007).

Úlcera cutánea en niño de 9 años

ingreso del paciente para iniciar tratamiento intravenoso con anfotericina B liposomal 165 mg/dia, que se mantiene durante 9 dias. La evolucion fue favorable. Los ensayos de PCR para Leishmania son positivos y, el estudio de las secuencias SSUrRNA e ITS-1 obtenidas con las distintas PCR indican que la infeccion esta causada por un parasito perteneciente al complejo Leishmania braziliensis. Descripcion clinica del caso