Evaluation of a new CT/NG/TV/MG Real-Time PCR Kit (Vircell) versus the Allplex STI Essential Assay (Seegene) for the diagnosis of sexually transmitted infections.

Sexually transmitted infections (STI) are a public health problem. Real-time PCR assays are the most sensitive test for screening and diagnosis of these infections. The aim of this study was to evaluate a new CT/NG/TV/MG Real-Time PCR (RT-PCR) kit (Vircell) for the detection of Chamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and Trichomonas vaginalis for the diagnosis of sexual transmitted infections using the Allplex STI Essential Assay (Seegene) as the reference's method. A total of 497 samples from different anatomical sites (endocervical, urethral, rectal, pharyngeal and urine) were analysed from October 2022 to February 2023. A total of 108 (21.73 %) and 106 (21.33 %) positive samples were found for any of the assays used. The most commonly detected pathogen was N. gonorrhoeae (52 samples; 10.46 %), and the least commonly detected was T. vaginalis (three samples; 0.60 %). The anatomical site with the highest prevalence of micro-organisms was a non-urogenital site, the pharynx (26 positive samples; 5.23 %). Using the Allplex STI Essential Assay (Seegene) as the reference method, the diagnosis performance showed that the average specificity of CT/NG/TV/MG RT-PCR Kit (Vircell) was 99.84 % and the sensitivity was 99.53 %. The overall concordance was k=0.98 (CI95 %; 0.96-1). In conclusion, the CT/NG/TV/MG RT-PCR Kit (Vircell) assay shows a good sensitivity and specificity and constitutes a promising and additional alternative to routine procedures for distinct types of clinical specimen in diagnosis STI.

Comparison of an Enzyme Linked-Immunosorbent Assay and a Chemiluminescent Immunoassay with an Immunofluorescence Assay for Detection of Phase II IgM and IgG Antibodies to Coxiella burnetii.

In this study, we have compared the detection of IgM and IgG against C. burnetii phase II of an enzyme-linked immunosorbent assay (ELISA) (Euroimmun) and a chemiluminescent immunoassay (CLIA) (VIRCLIA, Vircell). In addition, an indirect immunofluorescence assay (IFA) was used as a reference test. One hundred forty-eight sera were used for IgG evaluation, and eighty-eight for IgM. The sensitivity of ELISA and CLIA in detecting phase II IgM was excellent. On the other hand, the CLIA IgM showed better specificity than the ELISA IgM. As for phase II IgG, the specificity of ELISA and CLIA was similar, while the ELISA technique showed a higher sensitivity. In conclusion, the best system to detect phase II IgM antibodies against C. burnetii is the CLIA from Vircell, which is characterized by high sensitivity and specificity. For the detection of phase II IgG, the Euroimmun ELISA and Vircell CLIA assays are suitable for the determination of this marker in the laboratory, although the IgG ELISA has greater sensitivity.

In vitro activity of delafloxacin against anaerobic bacteria compared with other antimicrobials.

The aim of this study was to describe the in vitro activity of delafloxacin against 230 anaerobic isolates and compare it with the activity of other antimicrobials used against infections caused by anaerobic microorganisms. Minimal inhibitory concentrations (MICs) were lower for delafloxacin than for all other antibiotics tested with the exception of piperacillin-tazobactam and meropenem against Gram-positive anaerobic cocci. Only two (0.8 %) isolates of Bacteroides spp. showed a MIC ≥4 µg/mL. With some exceptions, the present results show lower MICs for delafloxacin in comparison to the other antibiotics used against anaerobes.

Otitis media tuberculosa: Presentación de un caso y revisión de la literatura europea / Tuberculous otitis media: A case presentation and review of european literature

La otitis media tuberculosa (OMT) es una afectación rara en nuestro medio que supone un reto en su diagnóstico debido a los síntomas inespecíficos que suele presentar. Este trabajo presenta nuestra experiencia en el diagnóstico de un caso de OMT en una mujer de 66 años con pérdida auditiva y otorrea crónica de más de 6 meses de evolución, que no respondía a los tratamientos convencionales. Además, se realiza una re visión de los casos publicados en los últimos 20 años (2000- 2022) en países de la Unión Europea (EU). Se incluyeron un total de 25 artículos con datos sobre 43 pacientes diagnostica dos de OMT. Las edades se situaron en un rango de: 3 meses - 87 años con un mayor porcentaje de mujeres (n=30; 69,77%). El tiempo medio de diagnóstico fue de 13,6 meses (rango, 1-72 meses). Los síntomas más comunes fueron otorrea (n=43; 100%), pérdida auditiva (n=37; 86,05%), perforación timpá nica (n=19; 44,18%), parálisis facial (n=12, 27,91%) y otalgia (n=13; 30,23%). La muestra empleada en mayor porcentaje para el diagnóstico fue la biopsia obtenida por mastoidectomía (n=34; 79,06%). Todos los pacientes fueron tratados con anti tuberculosos con una media de duración de 8,11 meses (rango, 6-12 meses). La secuela más frecuente fue la pérdida auditiva (n=28; 65,12%). La OMT debe incluirse en el diagnóstico dife rencial de las otitis supurativas crónicas ya que el diagnóstico y tratamiento precoz disminuyen la probabilidad de sufrir se cuelas irreversibles (AU)

Tuberculous otitis media (TOM) is a rare affectation in our environment that represents a challenge in its diagnosis due to the non-specific symptoms that it usually presents. This paper presents our experience in the diagnosis of a case of TOM in a 66-year-old woman with hearing loss and chronic otorrhea of more than 6 months of evolution that did not respond to con ventional treatments. In addition, a review of the cases pub lished in the last 20 years (2000-2022) in countries of the Eu ropean Union (EU) is carried out. The most common symptoms were otorrhea (n=43; 100%), hearing loss (n=37; 86.05%), eardrum perforation (n=19; 44.18%), facial paralysis (n=12, 27,91%) and ear pain (n=13; 30,23%). The most used sam ple for diagnosis was the biopsy obtained by mastoidectomy (n=34; 79.06%). All patients were given antituberculous ther apy for a mean duration of 8.11 months (range, 6-12 months). The most frequent aftereffect was hearing loss (n=28; 65.12%). TOM should be included in the differential diagnosis of chronic suppurative otitis, since early diagnosis and treatment reduce the probability of suffering irreversible sequelae (AU)

Differences between bloodstream infections involving Gram-positive and Gram-negative anaerobes.

BACKGROUND: The objectives of this study were to describe differences between bloodstream infections involving Gram-positive (GP) and Gram-negative (GN) anaerobic bacteria. METHODS: Patients with clinically significant anaerobic bacteremia detected between October 2016 and July 2022 in a tertiary hospital in Granada (Spain) were retrospectively included. Species identification was performed by MALDI-TOF MS and/or molecular methods. The association between variables was analyzed using contingency tables, applying the chi-square test when expected frequencies were adequate and the Fisher exact test when not. Variables were gathered at the time of the first positive blood culture. RESULTS: Out of 237 cases of anaerobic bloodstream infections detected, 127 (53.6%) were GN. Crude mortality was 20.3%, corresponding to 48 patients who died of causes directly attributable to bacteremia. The presence of malignant disease (p = 0.011), abdominal and/or pelvic surgery (p = 0.001), and transplantation (p = 0.008) were significantly associated with bacteremia due to GN bacteria, while the presence of diabetes mellitus was significantly associated with bacteremia due to GP bacteria (p = 0.022). The presence of both septic shock and mortality was more frequently associated with bacteremia due to GN versus GP bacteria. CONCLUSIONS: The association of certain variables with the presence of bloodstream infections due to GP or GN anaerobic bacteria may assist in selecting the optimal empirical therapeutic approach and improving the outcome of patients with these types of infection.

Neonatal invasive disease caused by Streptococcus agalactiae in Europe: the DEVANI multi-center study.

PURPOSE: Group B streptococcus (GBS) remains a leading cause of invasive disease, mainly sepsis and meningitis, in infants < 3 months of age and of mortality among neonates. This study, a major component of the European DEVANI project (Design of a Vaccine Against Neonatal Infections) describes clinical and important microbiological characteristics of neonatal GBS diseases. It quantifies the rate of antenatal screening and intrapartum antibiotic prophylaxis among cases and identifies risk factors associated with an adverse outcome. METHODS: Clinical and microbiological data from 153 invasive neonatal cases (82 early-onset [EOD], 71 late-onset disease [LOD] cases) were collected in eight European countries from mid-2008 to end-2010. RESULTS: Respiratory distress was the most frequent clinical sign at onset of EOD, while meningitis is found in > 30% of LOD. The study revealed that 59% of mothers of EOD cases had not received antenatal screening, whilst GBS was detected in 48.5% of screened cases. Meningitis was associated with an adverse outcome in LOD cases, while prematurity and the presence of cardiocirculatory symptoms were associated with an adverse outcome in EOD cases. Capsular-polysaccharide type III was the most frequent in both EOD and LOD cases with regional differences in the clonal complex distribution. CONCLUSIONS: Standardizing recommendations related to neonatal GBS disease and increasing compliance might improve clinical care and the prevention of GBS EOD. But even full adherence to antenatal screening would miss a relevant number of EOD cases, thus, the most promising prophylactic approach against GBS EOD and LOD would be a vaccine for maternal immunization.

Leishmaniasis vectors in the environment of treated leishmaniasis cases in Spain.

Transmission of leishmaniasis in endemic areas is characterized by microfocality related to the presence of the vector. Most entomological studies in southwestern Europe have focused on sylvatic areas and town outskirts, very few have sampled town or urban centres, and no survey has investigated inside households. The aim of this study was to determine the sand fly species diversity and vector density in the surroundings of human leishmaniasis cases compared with environments in which there was no association. Sand flies were captured in 26 households associated with recently treated leishmaniasis patients, 15 neighbouring houses without associated cases, and in others environments. Overall 7495 sand flies belonging to six species were captured. The highest sand fly density was found in farmhouses where there is a great availability of blood sources and breeding sites. In the environment of human leishmaniasis cases, Sergentomyia minuta was the most prevalent species followed by Phlebotomus perniciosus. Nevertheless, lower Leishmania infantum infection rates and lower intensity of infection were detected in S. minuta sand flies than in P. perniciosus. The density of P. perniciosus in households with recently treated leishmaniasis patients varies between 0 and 108 sand flies per light trap/night, with the maximum values corresponding to farmhouses. This species appears to be adapted to both indoors and outdoors domestic biotopes, including urban households.

HLA Class II Polymorphism and Humoral Immunity Induced by the SARS-CoV-2 mRNA-1273 Vaccine.

The vaccines designed against the SARS-CoV-2 coronavirus are based on the spike (S) protein. Processing of the S protein by antigen-presenting cells (APC) and its subsequent presentation to T cells is an essential part of the development of a humoral response. HLA-class II alleles are considered immune response genes because their codified molecules, expressed on the surface of APCs (macrophages, dendritic, and B cells) present antigenic peptides to T cell via their T cell receptor (TCR). The HLA-class II genes are highly polymorphic, regulating what specific peptides induce follicular helper T cells (TFH) and promote B lymphocyte differentiation into plasma or memory B cells. This work hypothesizes that the presence of certain HLA-class II alleles could be associated with the intensity of the humoral response (amount, length) to the SARS-CoV2 mRNA 1273 vaccine. We have studied the relationship between the HLA-class II typing of 87 health workers and the level of antibodies produced 30 days after vaccination. We show a possible association between the HLA-DRB1* 07:01 allele and the HLA-DRB1*07:01~DQA1*02:01~DQB1*02:02 haplotype to a higher production of antibodies 30 days after the administration of the second dose of mRNA-1273.

Misidentification of Phocaeicola (Bacteroides) dorei in two patients with bacteremia.

Phocaeicola (Bacteroides) dorei is a Gram-negative anaerobic bacillus that is rarely isolated from human specimens. Its accurate identification can be hampered by its close taxonomic relationship with Bacteroides vulgatus. We report on two patients with bacteremia due to P. (B.) dorei, which was initially identified as B. vulgatus by MALDI-TOF MS.

Study of humoral and cellular immunity in vaccinated with mRNA-1273.

The new vaccines against SARS-CoV-2 have raised a lot of expectations about their ability to induce immunity and the duration of this. This is the case of mRNA vaccines such as Moderna's mRNA-1273. Therefore, it is necessary to study the humoral and cellular immunity generated by these vaccines. Our objectives are determining what is the normal response of antibody production, and what is the level of protective antibodies and monitoring patients in case of subsequent infection with COVID-19. We present the first results of a longitudinal study of the humoral response in 601 health workers vaccinated with Moderna. The results show a humoral immunity at 90 days after the second dose of 100%, with a strong decrease between the levels of circulating anti-S IgG antibodies between days 30 and 90 post-vaccination. Observing a steeper decline in those who had higher titles at the beginning. In addition, we present a cellular response of 86% at three months after the second dose, which is related to low humoral response.

Comparative evaluation of chemiluminescent immunoassay and enzyme-linked immunosorbent assays for the diagnosis of West Nile virus infections.

In August 2020, anew West Nile virus (WNV) outbreak affected 71 people with meningoencephalitis in Andalusia (Spain). Samples from these individuals were received in our laboratory, a regional Virus Referral Centre. The aim of this study was to compare the agreement, sensitivity and specificity of findings between the WNV VIRCLIA IgG and IgM assay (Vircell, Spain) and the WNV ELISA IgM and IgG assay (Euroimmun, Germany) and to compare the performance of WNV VIRCLIA IgM and Euroimmun ELISA for cerebrospinal fluid (CSF) diagnosis. The study included 24 CSF samples (paired with serum samples) and 247 serum samples from 217 patients with suspected WNV infection (1 or 2 per patient). The agreement between ELISA and CLIA tests for IgM and Ig G detection in serum was 93% (kappa index = 0.85) and 96% (kappa index = 0.89) respectively. Sensitivity values of ELISA and CLIA tests for IgM in serum samples were 96.7% and 98.9%, respectively, and specificity values were 96.4% and 95.4% respectively. Sensitivity values of ELISA and CLIA test for IgG in serum samples were 91.1% and 97%, respectively, and specificity values were 100% and 98.8% respectively. Results obtained with ELISA and CLIA tests in CSF samples showed 75% agreement between them (kappa index = 0.51). According to these findings, the WNV VIRCLIA IgM and IgG monotest offers an accurate qualitative detection of WNV in serum and CSF specimens.

Leishmaniasis due to Leishmania infantum: Integration of human, animal and environmental data through a One Health approach.

The aim of this study was to explore Leishmania infantum epidemiology through a One Health approach that promotes a better estimation of leishmaniasis burden and a deeper understanding of the spatial distribution of the key actors of the parasite life cycle (vectors, reservoirs and humans). We conducted a 14-year mixed retrospective and prospective study of leishmaniasis cases in an endemic area in southern Spain (Granada province), to estimate the human incidence and its association with the vector presence, cryptic leishmaniasis rates and canine leishmaniasis prevalence. We found an annual linear increase in the incidence that cannot be fully explained by active case surveillance and the improvement of PCR diagnostic techniques. 49.4% of cases were not reported to the surveillance system. Approximately half of the human cases correspond to the visceral form that occurred more frequently in men; cutaneous, mucosal and cryptic forms were also detected. Leishmaniasis is no longer a disease of young children, accounting for a quarter of immunocompetent patients and most infected people remained asymptomatic. Human and canine leishmaniasis, cryptic or symptomatic, are present in the whole province, where there is a medium/high risk of the presence of Phlebotomus perniciosus, the main vector. We found association between the incidence of human leishmaniasis and the presence of the vector, but not with the prevalence of canine leishmaniasis and cryptic human leishmaniasis. A potential hot spot was also found, where high leishmaniasis incidence may be associated to the involvement of host species other than dogs.

A rare case of bacteremia caused by Propionibacterium (Propionimicrobium) lymphophilum.

Propionibacterium (Propionimicrobium) lymphophilum is a Gram-positive anaerobic rod involved in few human diseases. We report a rare case of bacteremia due to this microorganism in an elderly patient. A 95-year-old woman without a remarkable medical history presented with dyspnea, chest pain and fever for seven days. Blood cultures resulted in isolation of P. lymphophilum. Resistance only to metronidazole was found. Treatment with amoxicillin-clavulanic acid was established, and the patient was discharged and improvement of her general condition was documented.

Comparative evaluation of two immunoassays for serological diagnosis of hepatitis E.

Serological testing for immunoglobulin M (IgM) antibodies to Hepatitis E virus (HEV) is useful for diagnosing acute hepatitis E. Our aim was to evaluate the performance of chemiluminescence immunoassay (HEV IgM; Vircell, Granada, Spain) in comparison with a manual enzyme immunoassay (Anti-HEV virus; Wantai, Beijing, China) for anti-HEV IgM detection. Seventeen HEV-RNA positive and 69 HEV-RNA negative sera were tested by both assays. Virclia sensitivity was 94.1%, whereas that of the enzyme immunosorbent assay (EIA) was 76.5%; specificity was 98.5% and 97.1% for Virclia and EIA, respectively. Differences between both assays were not statistically significant. Virclia HEV IgM showed excellent performance and can be a good alternative to conventional enzyme immunoassays for the detection of IgM against HEV.

Antimicrobial susceptibility and clinical findings of significant anaerobic bacteria in southern Spain.

The objectives of this study were to report on the antimicrobial susceptibility of 276 clinically significant anaerobic bacteria belonging to the major genera isolated between May 2017 and November 2018 in a tertiary hospital in Granada (Spain) and to describe key clinical features of the patients. Species identification was performed by MALDI-TOF MS. Antimicrobial susceptibility tests were performed against penicillin, amoxicillin-clavulanic, imipenem, meropenem, moxifloxacin, clindamycin, metronidazole, vancomycin, and piperacillin-tazobactam using the gradient diffusion technique and EUCAST breakpoints (except for moxifloxacin). The most frequent anaerobes were Bacteroides (29.7%; n = 82), Clostridioides difficile (15.9%, n = 44), Prevotella (10.8%, n = 30), and Propionibacterium (10.7%, n = 25). Metronidazole was not universally active against all genera tested, and some isolates showed resistance to this drug. Almost all tested anaerobes were susceptible to carbapenems and amoxicillin-clavulanate except for Clostridioides difficile (resistance rate of 94%) and Bacteroides (19%), respectively. High overall resistance rates to clindamycin were observed, especially for genera Finegoldia (54%), Bacteroides (49%), and Prevotella (40%). Resistance rates to carbapenems and amoxicillin-clavulanate were very low for the majority of tested genera but were high for Clostridioides difficile and Bacteroides spp., respectively. Resistance to clindamycin was very high, especially for Bacteroides, Finegoldia magna, Prevotella and Peptoniphilus. Routine antimicrobial susceptibility testing for anaerobes contributes information on the global situation and allows empirical therapies to be selected in accordance with local data on resistant strains.

Corneal infection due to Moraxella nonliquefaciens / Infección corneal debida a Moraxella nonliquefaciens

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Maternal obesity and the risk of group B streptococcal colonisation in pregnant women.

The aim of the study was to test if maternal obesity and being overweight are independent risk factors for rectovaginal Group B Streptococcus (GBS) colonisation in pregnancy and for early onset GBS disease in the neonate. A case-control study of 9877 deliveries was conducted. The obese gravidas were significantly more likely to be colonised by GBS when compared with non-obese gravidas (22.7% versus 17.5%, P < .001). Obese gravidas were still 33% more likely than non-obese women to test positive for GBS after adjusting for the perinatal factors (adjusted OR 1.33 [95% CI 1.12-1.56]). The risk of early onset GBS disease was not calculated due to its very low incidence. The conclusion is that maternal obesity is a significant risk factor for GBS colonisation at term. Impact statement What is already known on this subject? Group B Streptococcus (GBS) is as an important cause of perinatal mortality and morbidity if prophylaxis is not performed. Intrapartum antibiotics are given if the carrier status is positive or unknown, provided that the risk factors are present. What do the results of this study add? Maternal obesity is a significant and independent risk factor for GBS colonisation at term. What are the implications of these findings for clinical practice and/or further research? Maternal obesity may be considered as a risk factor that should be taken into account in strategies for reducing GBS disease in neonates.

Detection of bacterial pathogens in sterile fluids with the FilmArray Meningitis/Encephalitis identification system / Detección de patógenos bacterianos en líquidos estériles con el sistema de identificación FilmArray Meningitis/Encefalitis

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