Transmitted drug resistance to antiretroviral drugs in Spain during the period 2019-2021.

To evaluate the prevalence of transmitted drug resistance (TDR) to nucleoside and nonnucleoside reverse transcriptase inhibitors (NRTI, NNRTI), protease inhibitors (PI), and integrase strand transfer inhibitors (INSTI) in Spain during the period 2019-2021, as well as to evaluate transmitted clinically relevant resistance (TCRR) to antiretroviral drugs. Reverse transcriptase (RT), protease (Pro), and Integrase (IN) sequences from 1824 PLWH (people living with HIV) were studied. To evaluate TDR we investigated the prevalence of surveillance drug resistance mutations (SDRM). To evaluate TCRR (any resistance level ≥ 3), and for HIV subtyping we used the Stanford v.9.4.1 HIVDB Algorithm and an in-depth phylogenetic analysis. The prevalence of NRTI SDRMs was 3.8% (95% CI, 2.8%-4.6%), 6.1% (95% CI, 5.0%-7.3%) for NNRTI, 0.9% (95% CI, 0.5%-1.4%) for PI, and 0.2% (95% CI, 0.0%-0.9%) for INSTI. The prevalence of TCRR to NRTI was 2.1% (95% CI, 1.5%-2.9%), 11.8% for NNRTI, (95% CI, 10.3%-13.5%), 0.2% (95% CI, 0.1%-0.6%) for PI, and 2.5% (95% CI, 1.5%-4.1%) for INSTI. Most of the patients were infected by subtype B (79.8%), while the majority of non-Bs were CRF02_AG (n = 109, 6%). The prevalence of INSTI and PI resistance in Spain during the period 2019-2021 is low, while NRTI resistance is moderate, and NNRTI resistance is the highest. Our results support the use of integrase inhibitors as first-line treatment in Spain. Our findings highlight the importance of ongoing surveillance of TDR to antiretroviral drugs in PLWH particularly with regard to first-line antiretroviral therapy.

Chemiluminescent Microparticle Immunoassay for the Diagnosis of Congenital Chagas Disease: A Prospective Study in Spain.

Congenital Chagas disease (CCD) has become a global health problem. Historically, the diagnosis of CCD has been carried out using parasitological methods and traditional serological techniques, however, new serological techniques such as chemiluminescent microparticle immunoassays (CMIA) have been developed in the last few years with many advantages compared with traditional serological tests. A total of 75 children born to 72 Latin American Chagas-infected mothers were consecutively enrolled and studied by CMIA and indirect immunofluorescence (IIF) at 0-2, 6, 9, and 12 months of age. At the end of the follow-up, 74 out of 75 children were considered uninfected and one child was diagnosed with CCD. Our study emphasizes the need to carry out serological follow-up on every newborn from a mother with Chagas disease and shows that CMIA assay is a great diagnostic tool as a single serological test at 9 months of age to rule out CCD or to identify possible transmission.

The algorithm used for the interpretation of doravirine transmitted drug resistance strongly influences clinical practice and guideline recommendations.

OBJECTIVES: We report the results of the reverse transcriptase (RT)/protease (PR) transmitted drug resistance (TDR) prevalence study in 2018, focusing on doravirine resistance-associated mutations and the differences observed when Stanford or French National Agency for AIDS Research (ANRS)/Spanish Network of AIDS Research (RIS)/IAS-USA resistance interpretation algorithms are used to describe clinically relevant resistance. METHODS: We used the WHO 2009 list to investigate the prevalence of NNRTI, NRTI and PI TDR, in treatment-naive HIV-1-infected patients, adding mutations E138A/G/K/Q/R, V106I, V108I, V179L, G190Q, H221Y, F227C/L/V, M230IDR, L234I, P236L and Y318F in RT. The prevalence of doravirine resistance-associated mutations, as described by Soulie et al. in 2019, was evaluated. Clinically relevant TDR was investigated using the latest versions of ANRS, RIS, IAS-USA and Stanford algorithms. RESULTS: NNRTI mutations were detected in 82 of 606 (13.5%) patients. We found 18 patients (3.0%) with NRTI mutations and 5 patients (0.8%) with PI mutations. We detected 11 patients harbouring doravirine resistance-associated mutations (prevalence of 1.8%). Furthermore, we observed important differences in clinically relevant resistance to doravirine when ANRS/RIS (0.7%), IAS-USA (0.5%) or Stanford algorithms (5.0%) were used. V106I, which was detected in 3.8% of the patients, was the main mutation driving these differences. V106I detection was not associated with any of the clinical, demographic or virological characteristics of the patients. CONCLUSIONS: The prevalence of NRTI and PI TDR remains constant in Spain. Doravirine TDR is very infrequent by RIS/ANRS/IAS-USA algorithms, in contrast with results using the Stanford algorithm. Further genotype-phenotype studies are necessary to elucidate the role of V106I in doravirine resistance.

Identification of Cryptosporidium subtype isolates from HIV-seropositive patients in Equatorial Guinea.

BACKGROUND: Cryptosporidium spp. are enteric parasites that infect humans and animals. In immunocompromised patients infection can be fatal. This study was conducted to identify sub-populations of Cryptosporidium hominis and C. parvum isolates from HIV-seropositive patients in Equatorial Guinea. METHODS: In a previous study conducted in Equatorial Guinea, faecal samples from 171 HIV patients with gastrointestinal symptoms were analyzed. Of these, 13 and 17 were positive for C. hominis and C. parvum, respectively. The isolates were characterized using gp60 gene analysis. RESULTS: The gp60 gene could only be detected in 57% (17/30) of cases (10 C. parvum and 7 C. hominis). Three C. hominis (Ia, Ib and Id) and two C. parvum (IIc and IIe) subtype families were detected, including several subtypes. CONCLUSIONS: The study identified a high diversity of Cryptosporidium subtypes, suggesting that anthroponotic transmission plays an important role in the epidemiology of Cryptosporidium spp. in HIV-seropositive patients in Equatorial Guinea.

Molecular characterization of Cryptosporidium isolates from humans in Equatorial Guinea.

The aim of the study was to perform a molecular characterization of clinical isolates of Cryptosporidium species from Equatorial Guinea. Standard laboratory methods were used to identify 35 cryptosporidiosis cases among 185 patients. PCR-RFLP successfully identified 34 Cryptosporidium species from these 35 cases, comprising C. parvum (52.9%), C. hominis (44.1%) and C. meleagridis (2.9%); over 90% of the species were isolated from HIV-positive patients. This is the first report of the molecular characterization of Cryptosporidium species isolated from humans in Equatorial Guinea and shows that zoonotic and anthroponotic transmission is present in this country.

A pilot study on the prevalenceof loiasis in Equatorial Guineausing an accurate Nested PCR

Background: In the last years loiasis has emerged as a public health problem in areas where Loa loa is co-endemic with Onchocerca volvulus, Wuchereria bancrofti and other filarial parasites. The objective of this work was to carried out a preliminary field study on the prevalence ofloiasis in Equatorial Guinea. Methods: The study design was carried out in three villages situated in the continental region and the insular region from Equatorial Guinea. A total of 236 human blood samples were obtained from individuals living in the continental region (n=142) and on the island of Bioko(n=96). Blood samples were diagnosed by leucoconcentration and microscopy examination formicrofilariae of L. loa and other filarial species. The molecular diagnosis was carried out by theL. loa specific nested PCR. Results: The study results shown a 22.8% of loiasis prevalence by microscopy observation, whereas the nested PCR revealed a prevalence of 76.4% in the continental region. In the 94samples obtained from individuals from the island of Bioko, loiasis was not detected either bymicroscopy analysis or by nested PCR. Conclusions: The nested PCR used in this work showed itself to be an accurate technique that detects the presence of L. loa DNA and it could be a useful complementary tool in ascertaining more precise estimates of the prevalence of loiasis in Equatorial Guinea (AU)

Fundamentos: En los últimos años la loasis se ha convertido en un grave problema de Salud Pública en áreas donde Loa loa es co-endémica con Onchocerca volvulus, Wuchereriabancrofti y otras filarias. El objetivo de este trabajo fue llevar a cabo un estudio preliminar de la prevalencia de loasis en Guinea Ecuatorial. Método: El diseño del estudio se llevo a cabo en tres poblaciones de la región continental yen la región insular de Guinea Ecuatorial. Se obtuvieron un total de 236 muestras de sangre procedentes de individuos de la región continental (n=142) y de la isla de Bioko (n=96). Las muestras fueron diagnosticadas por leuco concentración y examen microscópico de Loa loa y otras especies de filarias. El diagnóstico molecular se llevó a cabo mediante una nested PCR específica para Loa loa. Resultados: El resultado del estudio reveló una prevalencia de loasis del 22,8 % mediante diagnóstico microscópico y del 76,4% mediante nested PCR en la región continental. Loa loano fue detectada ni por análisis microscópico ni por nested PCR en las 94 muestras obtenidas de individuos de la Isla de Bioko. Conclusiones: La nested PCR utilizada en el trabajo y optimizada en nuestro laboratorio, es un método sensible que permite la detección de ADN de Loa loa y podría ser una herramienta complementaría útil en una evaluación más precisa de la prevalencia de loasis en Guinea Ecuatorial (AU)