In Vitro Antifungal Activity of Ibrexafungerp (SCY-078) Against Contemporary Blood Isolates From Medically Relevant Species of Candida: A European Study.

Background: Ibrexafungerp (SCY-078) is the newest oral and intravenous antifungal drug with broad activity, currently undergoing clinical trials for invasive candidiasis. Objective: The aim of this study was to assess the in vitro activity of ibrexafungerp and comparators against a collection of 434 European blood isolates of Candida. Methods: Ibrexafungerp, caspofungin, fluconazole, and micafungin minimum inhibitory concentrations (MICs) were collected from 12 European laboratories for 434 blood isolates, including 163 Candida albicans, 108 Candida parapsilosis, 60 Candida glabrata, 40 Candida tropicalis, 29 Candida krusei, 20 Candida orthopsilosis, 6 Candida guilliermondii, 2 Candida famata, 2 Candida lusitaniae, and 1 isolate each of Candida bracarensis, Candida catenulata, Candida dubliniensis, and Candida kefyr. MICs were determined by the EUCAST broth microdilution method, and isolates were classified according to recommended clinical breakpoints and epidemiological cutoffs. Additionally, 22 Candida auris from different clinical specimens were evaluated. Results: Ibrexafungerp MICs ranged from 0.016 to ≥8 mg/L. The lowest ibrexafungerp MICs were observed for C. albicans (geometric MIC 0.062 mg/L, MIC range 0.016-0.5 mg/L) and the highest ibrexafungerp MICs were observed for C. tropicalis (geometric MIC 0.517 mg/L, MIC range 0.06-≥8 mg/L). Modal MICs/MIC50s (mg/L) against Candida spp. were 0.125/0.06 for C. albicans, 0.5/0.5 for C. parapsilosis, 0.25/0.25 for C. glabrata, 0.5/0.5 for C. tropicalis, 1/1 for C. krusei, 4/2 for C. orthopsilosis, and 0.5/0.5 for C. auris. Ibrexafungerp showed activity against fluconazole- and echinocandin-resistant isolates. If adopting wild-type upper limits, a non-wild-type phenotype for ibrexafungerp was only observed for 16/434 (3.7%) isolates: 11 (4.6%) C. parapsilosis, 4 (5%) C. glabrata, and 1 (2.5%) C. tropicalis. Conclusion: Ibrexafungerp showed a potent in vitro activity against Candida.

Evaluation of 2 Commercial Assays for the Detection of Lymphogranuloma Venereum in Rectal Samples.

BACKGROUND: The early identification of the Chlamydia trachomatis variants that cause lymphogranuloma venereum (LGV) is very important to establish an adequate antibiotic treatment. This identification should be made with molecular techniques that are easy to perform and accessible to most microbiology laboratories. The objective of this study was to evaluate 2 real-time polymerase chain reaction (PCR)-based assay (VIASURE Haemophilus ducreyi + C. trachomatis (LGV) real-time PCR detection kit and the Allplex Genital ulcer Assay) for the detection of LGV in rectal samples. MATERIALS AND METHODS: Prospective study on positive rectal samples for C. trachomatis. All samples were processed in parallel by both tests. As a molecular reference method and to solve possible discrepancies between both assays, a PCR-based restriction fragment length polymorphism analysis of the major outer membrane protein gene (omp1) was used. RESULTS: In total, we detected 157 positive rectal samples for C. trachomatis, of which 36 were identified as LGV by PCR-based restriction fragment length polymorphism analysis. The positive percent agreement, negative percent agreement, and overall percent agreement were 88.9%, 100%, and 97.3%, respectively, for the Allplex Genital ulcer assay and 91.6%, 100%, and 97.1%, respectively, for the VIASURE assay. In the direct comparison between the Seegene assay and the VIASURE assay, we obtained a kappa concordance index of 0.98 between both tests. CONCLUSIONS: According to the results obtained, both tests could be used for the detection of LGV in rectal samples.

Updating of in vitro antifungal susceptibility tests. / Actualización de los métodos de estudio de sensibilidad in vitro a los antifúngicos.

Fungal diseases, including those caused by (multi)drug-resistant fungi, still represent a global public health concern. Information on the susceptibility of these microorganisms to antifungal agents must be quickly produced to help clinicians initiate appropriate antifungal therapies. Unfortunately, antifungal susceptibility tests are not as developed or widely implemented as antibacterial tests, being similar in design, accuracy and reproducibility, but also laborious and slow. In this article, we review the methods of in vitro susceptibility testing, both reference (CLSI and EUCAST), commercial and new methods based on proteomics (MALDI-TOF MS) and in the detection of resistance genes by nucleic acid amplification techniques. In addi-tion, we discuss the newly established clinical breakpoints, as well as the epidemiological cut-off points, which constitute a new category that can help in the early identification of isolates that have acquired resistance mechanisms. We also discuss the advantages and limitations of each of the methods studied. Therefore, we can conclude that, although there has been much progress in studies of in vitro susceptibility testing to antifungals, there are still limitations in its application in the daily routine of microbiology labo-ratories, although it seems that the future is promising with the new technologies based on proteomics and nucleic acid amplification. Supplement information: This article is part of a supplement entitled «SEIMC External Quality Control Programme. Year 2016¼, which is sponsored by Roche, Vircell Microbiologists, Abbott Molecular and Francisco Soria Melguizo, S.A. © 2019 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosasy Microbiología Clínica. All rights reserved.

Actualización de los métodos de estudio de sensibilidad in vitro a los antifúngicos / Updating of in vitro antifungal susceptibility tests

Las infecciones fúngicas, incluyendo aquellas producidas por hongos que pueden ser resistentes o multirresistentes a los antifúngicos, representan un serio problema de salud pública. La información sobre la sensibilidad de estos microorganismos a los distintos antifúngicos debe ser analizada lo más rápidamente posible para ayudar a los profesionales clínicos a instaurar un tratamiento adecuado. Desafortunadamente, las pruebas de sensibilidad a los antifúngicos no están tan desarrolladas ni implementadas como las de los antibacterianos, que son similares tanto en su diseño como en su precisión y reproducibilidad, pero laboriosas y lentas. En este artículo realizamos una revisión de los métodos de estudio de sensibilidad in vitro, tanto los de referencia (CLSI y EUCAST) como los comerciales y los nuevos métodos basados en la proteómica (MALDI-TOF MS) y en la detección de genes de resistencia por técnicas de amplificación de ácidos nucleicos. Además, se comentan los nuevos puntos de corte clínicos establecidos recientemente, así como los puntos de corte epidemiológicos, que se trata de una nueva categoría que puede ayudar a identificar de manera precoz las cepas aisladas que han adquirido mecanismos de resistencia. También se comentan las ventajas y las limitaciones de cada uno de los métodos revisados. Por tanto, puede concluirse que, aunque se ha avanzado mucho en los estudios de sensibilidad in vitro a los antifúngicos, aún existen limitaciones en su aplicación en la práctica diaria de un laboratorio de microbiología aunque parece que el futuro es esperanzador con las nuevas tecnologías basadas en la proteómica y en la amplificación de los ácidos nucleicos. Información sobre el suplemento: este artículo forma parte del suplemento titulado "Programa de Control de Calidad Externo SEIMC. Año 2016", que ha sido patrocinado por Roche, Vircell Microbiologists, Abbott Molecular y Francisco Soria Melguizo, S.A

Fungal diseases, including those caused by (multi)drug-resistant fungi, still represent a global public health concern. Information on the susceptibility of these microorganisms to antifungal agents must be quickly produced to help clinicians initiate appropriate antifungal therapies. Unfortunately, antifungal susceptibility tests are not as developed or widely implemented as antibacterial tests, being similar in design, accuracy and reproducibility, but also laborious and slow. In this article, we review the methods of in vitro susceptibility testing, both reference (CLSI and EUCAST), commercial and new methods based on proteomics (MALDI-TOF MS) and in the detection of resistance genes by nucleic acid amplification techniques. In addition, we discuss the newly established clinical breakpoints, as well as the epidemiological cut-off points, which constitute a new category that can help in the early identification of isolates that have acquired resistance mechanisms. We also discuss the advantages and limitations of each of the methods studied. Therefore, we can conclude that, although there has been much progress in studies of in vitro susceptibility testing to antifungals, there are still limitations in its application in the daily routine of microbiology labo-ratories, although it seems that the future is promising with the new technologies based on proteomics and nucleic acid amplification. Supplement information: This article is part of a supplement entitled "SEIMC External Quality Control Programme. Year 2016", which is sponsored by Roche, Vircell Microbiologists, Abbott Molecular and Francisco Soria Melguizo, S.A

A prospective survey of Aspergillus spp. in respiratory tract samples: Species identification and susceptibility patterns.

We analyzed the species distribution and susceptibility patterns of 433 strains of Aspergillus spp. isolated from respiratory samples of 419 in-patients included in multicenter prospective study (FUNGAE-IFI) between July 2014 and October 2015. Identification was carried out by conventional methods at each participating center and by molecular sequencing of a portion of the ß-tubulin gene at one of the centers. In vitro susceptibility was evaluated by broth microdilution methods and using the E-test (for cryptic species). Species identified included 249 A. fumigatus sensu stricto, 60 A. terreus sensu stricto, 47 A. flavus sensu stricto, 44 A. tubingensis, 18 A. niger sensu stricto , five A. nidulans sensu stricto, three A. tamarii, two A. calidoustus, two A. carneus, one A. acuelatus, one A. carbonarius, and one A. sydowii. Cryptic species were found in 12.5% of isolates (n = 54). The frequency of non-wild-type isolates for amphotericin B was 3.4% (n = 15) of the isolates tested and for azoles 3% (n = 10). None of the Aspergillus spp. were non-wild type to echinocandins. Of the 54 cryptic species only two strains were non-wild-type strains by microdilution method (3.7%) (two A. tubingensis, one to amphotericin B and another one to voriconazole) and by E-test method five strains of A. tubingensis showed high minimal inhibitory concentration (MIC) to amphotericin B (11.4%) and five to azoles (12.1%), one A. calidoustus strain showed high MICs for three azoles (50%), A. carneus to itraconazole (100%) and A. sydowii to amphotericin B and itraconazole (100%). These results provide relevant information on susceptibility patterns, frequency, and epidemiology of species involved in respiratory tract samples and of the incidence of recently described cryptic species.

Molecular Identification and Susceptibility Testing of Molds Isolated in a Prospective Surveillance of Triazole Resistance in Spain (FILPOP2 Study).

Antifungal resistance is increasing by the emergence of intrinsically resistant species and by the development of secondary resistance in susceptible species. A previous study performed in Spain revealed levels of azole resistance in molds of between 10 and 12.7%, but secondary resistance in Aspergillus fumigatus was not detected. We used itraconazole (ITZ)-supplemented medium to select resistant strains. A total of 500 plates supplemented with 2 mg/liter of ITZ were sent to 10 Spanish tertiary hospitals, and molecular identification and antifungal susceptibility testing were performed. In addition, the cyp51A gene in those A. fumigatus strains showing azole resistance was sequenced. A total of 493 isolates were included in the study. Sixteen strains were isolated from patients with an infection classified as proven, 104 were isolated from patients with an infection classified as probable, and 373 were isolated from patients with an infection classified as colonization. Aspergillus was the most frequent genus isolated, at 80.3%, followed by Scedosporium-Lomentospora (7.9%), Penicillium-Talaromyces (4.5%), Fusarium (2.6%), and the order Mucorales (1%). Antifungal resistance was detected in Scedosporium-Lomentospora species, Fusarium, Talaromyces, and Mucorales Three strains of A. fumigatus sensu stricto were resistant to azoles; two of them harbored the TR34+L98H mechanism of resistance, and the other one had no mutations in cyp51A The level of azole resistance in A. fumigatus remains low, but cryptic species represent over 10% of the isolates and have a broader but overall higher range of antifungal resistance.

Candida albicans Germ-Tube Antibody: Evaluation of a New Automatic Assay for Diagnosing Invasive Candidiasis in ICU Patients.

Testing for Candida albicans germ-tube antibody IFA IgG assay (CAGTA) is used to detect invasive candidiasis infection. However, most suitable assays lack automation and rapid single-sample testing. The CAGTA assay was adapted in an automatic monotest system (invasive candidiasis [CAGTA] VirClia® IgG monotest (VirClia®), a chemiluminescence assay with ready-to-use reagents that provides a rapid objective result. CAGTA assay was compared with the monotest automatic VirClia® assay in order to establish the diagnostic reliability, accuracy, and usefulness of this method. A prospective study with 361 samples from 179 non-neutropenic critically ill adults patients was conducted, including 21 patients with candidemia, 18 with intra-abdominal candidiasis, 84 with Candida spp. colonization, and 56 with culture-negative samples, as well as samples from ten healthy subjects. Overall agreement between the two assays (CAGTA and VirCLIA) was 85.3%. These assays were compared with the gold-standard method to determine the sensitivity, specificity as well as positive and negative predictive values. In patients with candidemia, values for CAGTA and VirCLIA assays were 76.2 versus 85.7%, 80.3 versus 75.8%, 55.2 versus 52.9%, and 91.4 versus 94.3%, respectively. The corresponding values in patients with intra-abdominal candidiasis were 61.1 versus 66.7%, 80.3 versus 75.8%, 45.8 versus 42.9%, and 88.3 versus 89.3%, respectively. No differences were found according to the species of Candida isolated in culture, except for Candida albicans and C. parapsilosis, for which VirClia® was better than CAGTA. According to these results, the automated VirClia® assay was a reliable, rapid, and very easy to perform technique as tool for the diagnosis invasive candidiasis.

Case Series Study of Invasive Pulmonary Aspergillosis.

Diagnosis of invasive pulmonary aspergillosis (IPA) is challenging. The objective of the study was to assess the value of microbiological tests to the diagnosis of IPA in the absence of non-specific radiological data. A retrospective study of 23 patients with suspicion of IPA and positivity of some microbiological diagnostic tests was performed. These tests included conventional microbiological culture, detection of Aspergillus galactomannan (GM) antigen and in some patients (1 â†’ 3)-ß-D-glucan (BDG) and Aspergillus fumigatus DNA using the LightCycler® SeptiFast test. In 10 patients with hematological malignancy, 6 cases were considered 'probable' and 4 'non-classifiable.' In 8 patients with chronic lung disease, 7 cases were classified as 'probable' and 1 as 'proven,' and in 5 patients with prolonged ICU stay (>7 days), there were 2 'proven' cases, 2 'non-classifiable' and 1 putative case. Microbiological culture was positive in 17 cases and 18 Aspergillus spp. were isolated (one mixed culture). A. fumigatus was the most frequent (44.4%) followed by A. tubingensis. The Aspergillus galactomannan (GM) antigen assay was positive in 21 cases (91.3%). The GM antigen and the (1 â†’ 3)-ß-D-glucan (BDG) assays were both performed in 12 cases (52.2%), being positive in 9. The SeptiFast test was performed in 7 patients, being positive in 4. In patients with non-classifiable pulmonary aspergillosis and one or more positive microbiological tests, radiological criteria may not be considered a limiting factor for the diagnosis of IPA.

Molecular identification, antifungal resistance and virulence of Cryptococcus neoformans and Cryptococcus deneoformans isolated in Seville, Spain.

Cryptococcal meningitis is one of the leading causes of death in HIV/AIDS patients. Our aim was to in order to characterise the epidemiology, antifungal susceptibility pattern and virulence of 28 Cyptococcus sp. strains recovered from 12 AIDS patients during two years in a Spanish single institution. Antifungal susceptibility testing was performed according to the CLSI protocols. Clinical strains were molecularly characterised by serotyping, mating type, PCR fingerprinting (M13 and GACA4 microsatellites) and analysis of two rDNA regions (IGS1 and ITS). Sequencing of the ERG11 gene was used to explore mechanisms of fluconazole resistance. Differences in virulence between species were studied in a Galleria mellonella infection model. Cryptococcus deneoformans and C. deneoformans x Cryptococcus neoformans hybrids were the most frequent variety (65%) followed by C. neoformans (35%). Strains were categorised according to 13 microsatellite genotypes and mixed infections could be detected in three patients. Twenty-nine per cent of the strains were fluconazole resistant. In one of the patients, the fluconazole resistance phenotype was associated with a point mutation in the ERG11 gene responsible for the amino acid substitution G470R. C. neoformans strains were able to kill G. mellonella larvae more efficiently than C. deneoformans and hybrids between both species. Precisely molecular characterisation of C. neoformans species is important for an accurate patient's management.

Epidemiología, factores de riesgo y sensibilidad in vitro en candidemias por especies diferentes de Candida albicans / Epidemiology, risk factors and in vitro susceptibility in candidaemia due to non-Candida albicans species

Antecedentes. La infección fúngica invasora ha aumentado en los últimos años por el incremento de los factores de riesgo; la candidemia es la principal manifestación clínica. Candida albicans es la especie más frecuente, aunque actualmente se ha observado un aumento en otras especies del género. Objetivos. Analizar la epidemiología, los factores de riesgo y la sensibilidad antifúngica de los aislamientos en hemocultivos de especies de Candida diferentes de C.albicans en nuestro hospital en los últimos 12años. Métodos. Se estudiaron retrospectivamente las características epidemiológicas de 107 pacientes con candidemia ingresados en nuestro hospital. Se determinó la sensibilidad de las especies de Candida al fluconazol, el itraconazol, el voriconazol, la anfotericinaB, la 5-fluorocitosina, la caspofungina, la micafungina y la anidulafungina mediante el método de microdilución Sensititre Yeast One (Izasa, España). Resultados. De los 109 aislamientos, 59 correspondieron a las siguientes especies de Candida (diferentes de C.albicans): 25 Candida parapsilosis complex, 14 Candida glabrata complex, 13 Candida tropicalis, 4 Candida krusei, una Candida lipolytica, una Candida membranaefaciens y una Candida pulcherrima. El factor de riesgo más frecuente en adultos y niños con candidemias por estas especies fue ser portador de catéter. El 8,5% de estos aislamientos fueron resistentes al fluconazol. Conclusiones. El resultado de nuestro trabajo confirma la necesidad de conocer la epidemiología de las especies de Candida diferentes de C.albicans, su sensibilidad in vitro y los factores de riesgo asociados, especialmente en pacientes con dichos factores (AU)

Background. Invasive fungal infection (IFI) has increased in recent years due to there being a greater number of risk factors. IFI caused by Candida is the most frequent, and although Candida albicans is the most isolated species, there is currently a decrease of C. albicans and an increase of other species of the genus. Aims. To analyse the epidemiology, risk factors, and antifungal susceptibility of blood culture isolates of non-C.albicans Candida species in our hospital in the last 12years. Methods. A retrospective study was conducted on 107 patients with candidaemia admitted to our hospital. Candida isolates susceptibility to fluconazole, itraconazole, voriconazole, amphotericinB, 5-fluorocytosine, caspofungin, micafungin, and anidulafungin was determined by means of a microdilution technique (Sensititre Yeast One; Izasa, Spain). Results. From a total of 109 strains, 59 belonged to non-C. albicans Candida species: 25 Candida parapsilosis complex, 14 Candida glabrata complex, 13 Candida tropicalis, 4 Candida krusei, 1 Candida lipolytica, 1 Candida membranaefaciens, and 1 Candida pulcherrima. The most common risk factor in adults and children was catheter use. It was observed that 8.5% of those non-C.albicans strains were resistant to fluconazole. Conclusions. The results of this work confirm that it is necessary to know the epidemiology of non-C.albicans Candida species, the in vitro susceptibility of the species involved, and the main risk factors, especially in patients with predisposing conditions (AU)

[Epidemiology, risk factors and in vitro susceptibility in candidaemia due to non-Candida albicans species]. / Epidemiología, factores de riesgo y sensibilidad in vitro en candidemias por especies diferentes de Candida albicans.

BACKGROUND: Invasive fungal infection (IFI) has increased in recent years due to there being a greater number of risk factors. IFI caused by Candida is the most frequent, and although Candida albicans is the most isolated species, there is currently a decrease of C. albicans and an increase of other species of the genus. AIMS: To analyse the epidemiology, risk factors, and antifungal susceptibility of blood culture isolates of non-C.albicans Candida species in our hospital in the last 12years. METHODS: A retrospective study was conducted on 107 patients with candidaemia admitted to our hospital. Candida isolates susceptibility to fluconazole, itraconazole, voriconazole, amphotericinB, 5-fluorocytosine, caspofungin, micafungin, and anidulafungin was determined by means of a microdilution technique (Sensititre Yeast One; Izasa, Spain). RESULTS: From a total of 109 strains, 59 belonged to non-C. albicans Candida species: 25 Candida parapsilosis complex, 14 Candida glabrata complex, 13 Candida tropicalis, 4 Candida krusei, 1 Candida lipolytica, 1 Candida membranaefaciens, and 1 Candida pulcherrima. The most common risk factor in adults and children was catheter use. It was observed that 8.5% of those non-C.albicans strains were resistant to fluconazole. CONCLUSIONS: The results of this work confirm that it is necessary to know the epidemiology of non-C.albicans Candida species, the in vitro susceptibility of the species involved, and the main risk factors, especially in patients with predisposing conditions.

Contribution of Candida biomarkers and DNA detection for the diagnosis of invasive candidiasis in ICU patients with severe abdominal conditions.

BACKGROUND: To assess the performance of Candida albicans germ tube antibody (CAGTA), (1 → 3)-ß-D-glucan (BDG), mannan antigen (mannan-Ag), anti-mannan antibodies (mannan-Ab), and Candida DNA for diagnosing invasive candidiasis (IC) in ICU patients with severe abdominal conditions (SAC). METHODS: A prospective study of 233 non-neutropenic patients with SAC on ICU admission and expected stay ≥ 7 days. CAGTA (cutoff positivity ≥ 1/160), BDG (≥80, 100 and 200 pg/mL), mannan-Ag (≥60 pg/mL), mannan-Ab (≥10 UA/mL) were measured twice a week, and Candida DNA only in patients treated with systemic antifungals. IC diagnosis required positivities of two biomarkers in a single sample or positivities of any biomarker in two consecutive samples. Patients were classified as neither colonized nor infected (n = 48), Candida spp. colonization (n = 154) (low-grade, n = 130; high-grade, n = 24), and IC (n = 31) (intra-abdominal candidiasis, n = 20; candidemia, n = 11). RESULTS: The combination of CAGTA and BDG positivities in a single sample or at least one of the two biomarkers positive in two consecutive samples showed 90.3 % (95 % CI 74.2-98.0) sensitivity, 42.1 % (95 % CI 35.2-98.8) specificity, and 96.6 % (95 % CI 90.5-98.8) negative predictive value. BDG positivities in two consecutive samples had 76.7 % (95 % CI 57.7-90.1) sensitivity and 57.2 % (95 % CI 49.9-64.3) specificity. Mannan-Ag, mannan-Ab, and Candida DNA individually or combined showed a low discriminating capacity. CONCLUSIONS: Positive Candida albicans germ tube antibody and (1 → 3)-ß-D-glucan in a single blood sample or (1 → 3)-ß-D-glucan positivity in two consecutive blood samples allowed discriminating invasive candidiasis from Candida spp. colonization in critically ill patients with severe abdominal conditions. These findings may be helpful to tailor empirical antifungal therapy in this patient population.

Performance of the HSV OligoGen kit for the diagnosis of herpes simplex virus type 1 and 2.

PCR methods are nowadays between the most rapid and sensitive methods for screening and diagnosing herpes simplex virus (HSV) type 1 and 2. The aim of this study was to analyze the reliability, accuracy, and usefulness of the new assay HSV OligoGen kit in comparison with the Roche LightCycler HSV ½ Qual Kit assay for the detection of HSV in clinical samples. For this analysis, a prospective study was designed for detection of HSV-1 and HSV-2 including 110 ulcer specimens, 48 urine, 48 endocervical, 43 cerebral spinal fluids, 4 urethral and 3 pharyngeal swabs that were sent from a regional STI clinic or an Intensive Clinical Unit, both in Seville, Spain. In comparison to the Roche LightCycler HSV ½ Qual Kit assay, sensitivity, specificity, positive and negative predicative values, and kappa value for HSV detection using the HSV OligoGen kit were 96.2%, 100%, 100%, 98.3%, and 0.97 for HSV-1, respectively. For HSV-2, the corresponding values were 98.3%, 100%, 100%, 99.5%, and 0.98, respectively. Statistical data obtained in this study confirms the usefulness and reliable results of this new assay.

Comparison of the CT OligoGen kit with cobas 4800 assay for detection of Chlamydia trachomatis / Comparación del kit CT OligoGen con el ensayo Cobas 4800 para la detección de Chlamydia trachomatis

INTRODUCTION: A prospective study was designed to assess the performance of the new CT OligoGen kit and the cobas 4800 assay for detection of Chlamydia trachomatis. METHODS: A set of samples that included urine samples (n = 212), endocervical (n = 167), rectal (n = 53), pharyngeal (n = 7) and urethral swabs (n = 3). The samples were sent from a regional sexually transmitted diseases (STD) clinic in Seville, Spain, and were collected from 261 men and 181 women. Discordant results were re-analyzed and clinical data and other tests were reviewed in order to resolve them. RESULTS: Sensitivity, specificity, positive predicative value (PPV), negative predictive value (NPV) and kappa value for C. trachomatis detection using the CT OligoGen kit were 98.5%, 100%, 100%, 95.4% and 0.97, respectively. Conclusions This new kit had a high sensitivity, specificity, PPV and NPV for C. trachomatis, therefore the performance profile confirms the usefulness and reliable results of this new assay

INTRODUCCIÓN: Se diseñó un estudio prospectivo para evaluar las características del nuevo kit CT OligoGen en comparación con el test cobas 4800 para la detección de Chlamydia trachomatis. MÉTODOS: Se analizaron una serie de muestras que incluían orinas (n = 212), exudados endocervicales (n = 167), rectales (n = 53), faríngeos (n = 7) y uretrales (n = 3). Estas muestras provenían de un centro de infecciones de transmisión sexual (Sevilla) y pertenecían a 261 hombres y 181 mujeres. Los resultados discordantes se reanalizaron y revisaron historias clínicas y otras pruebas para resolverlas. RESULTADOS: Los valores de sensibilidad, especificidad, valor predictivo positivo (VPP) y negativo (VPN) y valor kappa para el kit CT OligoGen fue 98,5%, 100%, 100%, 95,4% and 0,97, respectivamente. CONCLUSIONES: Este nuevo kit tuvo una alta sensibilidad, especificidad, VPP y VPN para la detección de C. trachomatis, por lo que esta evaluación confirma su utilidad y fiabilidad

ß-D-Glucan and Candida albicans germ tube antibody in ICU patients with invasive candidiasis.

PURPOSE: To assess the performance of (1→3)-ß-D-glucan (BDG) and Candida albicans germ tube antibody (CAGTA) for the diagnosis of invasive candidiasis (IC) in a prospective cohort of 107 unselected, non-neutropenic ICU patients. METHODS: BDG (cutoff positivity ≥80 pg/mL) and CAGTA (cutoff positivity ≥1/160) assays were performed twice a week. Confounding factors included amoxicillin-clavulanate and piperacillin-tazobactam treatments, recent surgery, Gram-positive bloodstream infection, renal replacement therapy, and enteral nutrition. Patients were classified as neither colonized nor infected (n = 29), Candida spp. colonization (n = 63) (low grade, n = 32; high grade, n = 31), and invasive candidiasis (IC) (n = 15). RESULTS: BDG levels were higher in patients with IC and high-grade colonization than in the remaining groups (p = 0.012), and two consecutive measurements ≥80 pg/mL discriminated IC from the remaining groups (sensitivity 80%, specificity 75.7%). For the discrimination between IC and Candida spp. colonization, the AUC for the maximum value of BDG was 0.667 (95% CI 0.544-0.790) and for the maximum value of CAGTA 0.545 (95% CI 0.395-0.694). Significant changes of BDG and CAGTA kinetics in IC patients treated with antifungals were not observed. In patients neither colonized nor infected or with low-grade Candida spp. colonization, none of the confounding factors was associated with a significant increase in BDG positivity. CONCLUSIONS: Two consecutive BDG levels ≥80 pg/mL allowed discrimination among IC and high-grade colonization. Systemic antifungal therapy could not be monitored with biomarker kinetics, and BDG levels were not subject to interference by confounding factors in either colonized or infected patients or with low-grade colonization.

Comparison of the CT OligoGen kit with cobas 4800 assay for detection of Chlamydia trachomatis.

INTRODUCTION: A prospective study was designed to assess the performance of the new CT OligoGen kit and the cobas 4800 assay for detection of Chlamydia trachomatis. METHODS: A set of samples that included urine samples (n=212), endocervical (n=167), rectal (n=53), pharyngeal (n=7) and urethral swabs (n=3). The samples were sent from a regional sexually transmitted diseases (STD) clinic in Seville, Spain, and were collected from 261 men and 181 women. Discordant results were re-analyzed and clinical data and other tests were reviewed in order to resolve them. RESULTS: Sensitivity, specificity, positive predicative value (PPV), negative predictive value (NPV) and kappa value for C. trachomatis detection using the CT OligoGen kit were 98.5%, 100%, 100%, 95.4% and 0.97, respectively. CONCLUSIONS: This new kit had a high sensitivity, specificity, PPV and NPV for C. trachomatis, therefore the performance profile confirms the usefulness and reliable results of this new assay.

Comparison of urine and cervical samples for detecting human papillomavirus (HPV) with the Cobas 4800 HPV test.

BACKGROUND: Human papillomavirus (HPV) is the main cause of cervical cancer. The development of non-invasive self-sample collection methods would have the potential advantage of increasing the acceptance of the screening procedures. OBJECTIVES: To compare human papillomavirus (HPV) DNA detection and genotyping with the Cobas 4800 HPV test (Roche Diagnostic, Spain) on paired cervical and first voided urine. STUDY DESIGN: Paired urine and cervical samples were collected from 125 women referred for evaluation of abnormal Pap smear results. RESULTS: The overall percent agreement between HPV detection in urine and cervical samples was 88%. A substantial concordance rate of HPV DNA detection in both samples was observed (κ=0.76; 95% IC: 64-87). In this high prevalence population the sensitivity, specificity, NPV and PPV for detection of HPV DNA from urine versus cervical samples were 90.5% (95% IC: 80-95%), 85%, (95% IC: 74-92%), 89.8% (95% IC: 79.5-95.3) and 86.4% (95% IC: 76.1-92.7) respectively. Compared to histologically confirmed CIN 2/3 disease, the clinical sensitivity and specificity for the detection of high-risk HPV in urine samples were 95% (95% IC: 76-97%) and 52.4% (95% IC: 40-64%) respectively. CONCLUSIONS: These results suggest that urine samples processed with Cobas 4800 HPV test may be useful for clinical management of HPV infection.

Métodos microbiológicos para el diagnóstico, manejo y estudio de la infección fúngica invasora / Microbiological procedures for the diagnosis, management, and study of invasive fungal infections

Las infecciones fúngicas invasoras (IFI) son infecciones de difícil diagnóstico y causan una alta mortalidad a una amplia variedad de enfermos. Debido a las limitaciones del cultivo microbiológico, se han desarrollado otras técnicas diagnósticas, entre las que destacan la determinación sérica de galactomanano o beta-1,3-d-glucano, y los anticuerpos antimicelio y antimanano. La aplicación de la biología molecular y la espectrometría de masas (MALDI-TOF) son esperanzadoras para la optimización del diagnóstico y el manejo de pacientes con IFI. En lo relativo a los fármacos antifúngicos, el desarrollo de nuevos puntos de corte especie-específicos para clasificar a los aislados como sensibles o resistentes, y la necesidad de monitorización sérica de azoles, merecen especial atención

Invasive fungal infections (IFIs) are difficult to diagnose and cause a high mortality to an expanding spectrum of patients. Culture of clinical samples has limitations for the diagnosis of IFI and alternative procedures have been developed. Among them, serum determination of galactomannan or beta-1,3-d-glucan, and antimicelium and antimannan antibodies are relevant. The use of molecular procedures and mass spectrometry (MALDI-TOF) are encouraging tools for the optimization of the diagnosis and management of patients with IFI. The proposal of species-specific breakpoint to classify the isolates as resistant or susceptible to antifungal agents and the necessity of monitor the azole serum levels deserve greater attention

Impact of therapeutic strategies on the prognosis of candidemia in the ICU.

OBJECTIVES: To determine the epidemiology of Candida bloodstream infections, variables influencing mortality, and antifungal resistance rates in ICUs in Spain. DESIGN: Prospective, observational, multicenter population-based study. SETTING: Medical and surgical ICUs in 29 hospitals distributed throughout five metropolitan areas of Spain. PATIENTS: Adult patients (≥ 18 yr) with an episode of Candida bloodstream infection during admission to any surveillance area ICU from May 2010 to April 2011. INTERVENTIONS: Candida isolates were sent to a reference laboratory for species identification by DNA sequencing and susceptibility testing using the methods and breakpoint criteria promulgated by the European Committee on Antimicrobial Susceptibility Testing. Prognostic factors associated with early (0-7 d) and late (8-30 d) mortality were analyzed using logistic regression modeling. MEASUREMENTS AND MAIN RESULTS: We detected 773 cases of candidemia, 752 of which were included in the overall cohort. Among these, 168 (22.3%) occurred in adult ICU patients. The rank order of Candida isolates was as follows: Candida albicans (52%), Candida parapsilosis (23.7%), Candida glabrata (12.7%), Candida tropicalis (5.8%), Candida krusei (4%), and others (1.8%). Overall susceptibility to fluconazole was 79.2%. Cumulative mortality at 7 and 30 days after the first episode of candidemia was 16.5% and 47%, respectively. Multivariate analysis showed that early appropriate antifungal treatment and catheter removal (odds ratio, 0.27; 95% CI, 0.08-0.91), Acute Physiology and Chronic Health Evaluation II score (odds ratio, 1.11; 95% CI, 1.04-1.19), and abdominal source (odds ratio, 8.15; 95% CI, 1.75-37.93) were independently associated with early mortality. Determinants of late mortality were age (odds ratio, 1.04; 95% CI, 1.01-1.07), intubation (odds ratio, 7.24; 95% CI, 2.24-23.40), renal replacement therapy (odds ratio, 6.12; 95% CI, 2.24-16.73), and primary source (odds ratio, 2.51; 95% CI, 1.06-5.95). CONCLUSIONS: Candidemia in ICU patients is caused by non-albicans species in 48% of cases, C. parapsilosis being the most common among these. Overall mortality remains high and mainly related with host factors. Prompt adequate antifungal treatment and catheter removal could be critical to decrease early mortality.