Evaluación de la prueba Aspergillus Galactomannan Ag VircliaR Monotest como alternativa a kit Platelia ™ Aspergillus EIA / Evaluation of the Aspergillus Galactomannan ag VircliaR Monotest test as an alternative to Platelia™ Aspergillus EIA kit

INTRODUCCIÓN: La prueba Aspergillus galactomannan Ag Virclia® (GM-VClia) es una técnica de galactomanano monotest, auto-matizada, basada en inmunoensayo quimioluminiscente (CLIA). OBJETIVO: Evaluar el desempeño del test de GM-VClia en muestras de suero y lavado bronquioalveolar (LBA) procesadas previamente con el kit Platelia™ Aspergillus EIA (GM-Plat). MATERIALES Y MÉTODOS: Se estudiaron 56 muestras de suero y 40 de LBA, correspondientes a un total de 59 pacientes (algunos con determinación de galactomamano en ambas muestras) con enfermedades pulmonares, hematológicas, LES, Covid-19 y tumores, entre otros. Trece pacientes tuvieron aspergilosis invasora (1 probada y 12 probables). RESULTADOS: La correlación entre ambos métodos para suero y LBA fue r = 0,8861 p < 0,0001 y r = 0,6368 p < 0,001, respectivamente. Hubo una concordancia global de 67,7% (65/96), siendo de 85,7% (48/56) en sueros y 42,5,0% (14/49) en LBA. Al subir el punto de corte en LBA por GM-VClia la concordancia aumentó a 85,7%. CONCLUSIONES: Se observó una mayor correlación y concordancia en sueros que en LBA. El kit GM-VClia presentó una mayor sensibilidad y valor predictor negativo (VPN), que el kit GM-Plat. Las desventajas de GM-VClia, la constituyen la categoría "dudoso", que dificulta la interpretación y que, con los puntos de corte actuales en LBA, la correlación con GM-Plat es menor. Las ventajas son su mayor sensibilidad, facilidad de procesamiento y una mayor rapidez en los resultados.

BACKGROUND: The Aspergillus Galactomannan Ag Virclia® (GMVClia) test is a monotest and automated galactomannan technique based on chemiluminescent immunoassay (CLIA). AIM: To evaluate the performance of the GM-VClia test in serum and bronchioalveolar lavage (BAL) samples previously processed with the Platelia ™ Aspergillus EIA kit (GM-Plat). METHODS: 56 samples of serum 40 from BAL (some of them with galactomaman determination in both samples), from patients with pulmonary diseases, hematological diseases, SLE, Covid-19 and tumors, among others, were studied. Thirteen patients had invasive aspergillosis (1 proven and 12 probable). RESULTS: The correlation between both methods for serum and BAL was r = 0.8861 p < 0.0001 and r = 0.6368 p < 0.001, respectively. There was a global concordance of 67.7% (65/96), being 85.7% (48/56) in sera and 42.5.0% (14/49) in BAL. By raising the cut-off point in LBA by GM-VClia, the agreement increased to 85.7%. CONCLUSION: A greater correlation and concordance was observed in sera than in BAL. The GM-VClia kit had a higher sensitivity and NPV than the GM-Plat kit. The disadvantages of GM-VClia are the "doubtful" category, which makes interpretation difficult and that with the current cut-off points in LBA the correlation with GM-Plat is lower. The advantages are its greater sensitivity, ease of processing and faster results.

Aumento en la positividad del biomarcador galactomanano durante la pandemia de COVID-19 en la Quinta Región: Valparaíso / Increase in the positivity of the galactomannan biomarker during the COVID-19 pandemic in the Fifth Region: Valparaíso

INTRODUCCIÓN: La actual pandemia provocada por SARS-CoV-2 ha provocado una alta carga en la salud pública y privada. Se han descrito casos y series de aspergilosis invasora asociada a pacientes con COVID-19 en ventilación mecánica. OBJETIVO: Describir el aumento en la positividad del biomarcador galactomanano (GM) durante la pandemia de COVID-19 en la Quinta Región: Valparaíso. MATERIALES Y MÉTODO: Estudio descriptivo, retrospectivo. Se revisó la cantidad y los resultados de GM, tanto de lavado bronco-alveolar (LBA) como en suero y los cultivos de LBA enviados al laboratorio de Micología de la Universidad de Valparaíso, desde enero y hasta septiembre del año 2020; luego se compararon con los exámenes recibidos en el mismo período del año 2019. RESULTADOS: Se observó un aumento significativo de los GM realizados en LBA, concentrándose principalmente entre los meses de julio y septiembre. El 29% de las muestras del año 2020 tenía el antecedente de ser de pacientes con COVID-19. Del total de muestras positivas durante el año de la pandemia, 5/12 fueron en pacientes con COVID-19. CONCLUSIONES: Hubo un aumento significativo de los GM realizados en LBA durante la pandemia, concentrándose principalmente entre los meses de julio-septiembre.

BACKGROUND: The current pandemic due to SARS-CoV-2 has caused a high burden on health. Cases and series of invasive aspergillosis associated with COVID-19 patients (CAPA) on mechanical ventilation have been described. AIM: To describe the increase in the positivity of the galactomannan (GM) biomarker during the COVID-19 pandemic in the Fifth Region: Valparaíso. METHOD: Retrospective descriptive study. The GM results in both broncho-alveolar lavage (BAL) and serum and the BAL cultures that were sent to the Mycology Laboratory of the University of Valparaíso from January to September 2020 were reviewed; then they were compared with the examinations of the same period of 2019. RESULTS: There was a significant increase in GMs carried out in LBA during the pandemic, concentrating mainly between the months of July-September. CONCLUSIONS: There was a significant increase in GM carried out in LBA during the pandemic, concentrating mainly between the months of July-September.

Enzymatic preparation of manno-oligosaccharides from locust bean gum and palm kernel cake, and investigations into its prebiotic activity

BACKGROUND: Manno-oligosaccharides (MOS) is known as a kind of prebiotics. Mannanase plays a key role for the degradation of mannan to produce MOS. In this study, the mannanases of glycoside hydrolase (GH) families 5 Man5HJ14 and GH26 ManAJB13 were employed to prepare MOS from locust bean gum (LBG) and palm kernel cake (PKC). The prebiotic activity and utilization of MOS were assessed in vitro using the probiotic Lactobacillus plantarum strain. RESULTS: Galactomannan from LBG was converted to MOS ranging in size from mannose up to mannoheptose by Man5HJ14 and ManAJB13. Mannoheptose was got from the hydrolysates produced by Man5HJ14, which mannohexaose was obtained from LBG hydrolyzed by ManAJB13. However, the same components of MOS ranging in size from mannose up to mannotetrose were observed between PKC hydrolyzed by the mannanases mentioned above. MOS stability was not affected by high-temperature and high-pressure condition at their natural pH. Based on in vitro growth study, all MOS from LBG and PKC was effective in promoting the growth of L. plantarum CICC 24202, with the strain preferring to use mannose to mannotriose, rather than above mannotetrose. CONCLUSIONS: The effect of mannanases and mannan difference on MOS composition was studied. All of MOS hydrolysates showed the stability in adversity condition and prebiotic activity of L. plantarum, which would have potential application in the biotechnological applications.

Applicability of the Platelia EIA® Aspergillus test for the diagnosis of aspergilosis in penguins / Aplicabilidade do Platelia EIA® Aspergillus como teste diagnostico da aspergilose em pinguins

Abstract Even today, an effective diagnostic test for aspergillosis in penguins is unknown, being the gold standard post-mortem examinations. The fungal antigen galactomannan (GM) has been used as a biomarker of disease in humans and is detected by the Platelia Aspergillus EIA (BioRad)®, a commercial kit based on the sandwich ELISA technique. It is standardized for use in neutropenic patients, however studies have demonstrated its usefulness also possible for birds. The aim of our study was to evaluate the effectiveness of Platelia Aspergillus EIA® test (BioRad-US) in the diagnosis of aspergillosis in Magellanic penguins, determining sensitivity, specificity, and positive and negative predictive values for different cut-off points. Were included in the study, blood serum samples (n = 29) Magellanic penguins in captivity that died by aspergillosis. Detection of GM was performed following manufacturer's instructions and the GM index was obtained by dividing the average value of OD of the duplicate of the clinical sample by duplicate OD of the average value of the cut-off sample provided by the kit. Through information database results were obtained for the presence of anti-Aspergillus fumigatus antibodies detected by agar gel immunodiffusion (AGID) for all serum samples. Results were analyzed using chi-square test and Kruskal-Wallis from SPSS 20.0, IBM®. ROC curve was obtained and from this, rates of sensitivity, specificity, positive and negative predictive values were also calculated based on four different cutoff points (0.5, 1.0, 1.5 and 2.0). The serum GM index did not differ between animals of the case and control group (pkw =0.097). In determining the ROC curve for serum GM detection the value of area under the curve was 0.635. From the values ​​determined by the coordinate of the curve, four different cut points (0.5, 1.0, 1.5 and 2.0) were analyzed, resulting in sensitivity rates ranging from 86.2 to 34.5% % and specificity between 87% and 26.1%. By comparing the serum GM index in group case as the presence or absence of antibodies detected by AGID was found p=0.503. The detection of GM the Platelia Aspergillus EIA® test seems is not be useful for the diagnosis of aspergillosis in naturally infected penguins.

Resumo Ainda hoje, um teste diagnóstico eficaz para aspergilose em pinguins não é conhecido, sendo o padrão-ouro os exames post-mortem. O antígeno fúngico galactomanana (GM) tem sido utilizado como biomarcador da doença em humanos, sendo detectado pelo Platelia Aspergillus EIA (BioRad)®, um kit comercial que se baseia na técnica ELISA sanduíche. É padronizado para utilização em pacientes neutropênicos, no entanto estudos tem demonstrado sua possível utilidade também para aves.O objetivo de nosso estudo foi avaliar a eficácia do teste Platelia Aspergillus EIA® (BioRad-US) no diagnóstico da aspergilose em pinguins-de-Magalhães, determinando sensibilidade, especificidade e valores preditivos positivos e negativos em diferentes pontos de corte. Foram incluídas no estudo, amostras de soro sanguíneo (n=29) de pinguins-de-Magalhães em cativeiro que vieram a óbito por aspergilose. A detecção de GM foi realizada seguindo instruções do fabricante e o índice de GM foi obtido dividindo o valor da média da DO da duplicata da amostra clínica pelo valor da média da DO da duplicata da amostra de cut-off fornecida pelo kit. Através de informações em banco de dados foram obtidos resultados sobre a presença de anticorpos anti-Aspergillus fumigatus, detectada por Imunodifusão em gel de ágar (IDGA) em todas as amostras séricas. Os resultados foram analisados utilizando-se teste de qui-quadrado e Kruskal-Wallis a partir do programa estatístico SPSS 20.0, IBM®. Curva ROC foi obtida e a partir desta, taxas de sensibilidade, especificidade, valores preditivo positivo e negativo foram igualmente calculados considerando quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0). O índice de GM sérica não diferiu entre os animais do grupo caso e controle (pKW = 0.097). Na determinação da curva ROC para detecção de GM sérica o valor da área sobre a curva foi de 0.635. A partir dos valores determinados pelas coordenadas da curva, quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0) foram analisados, resultando em taxas de sensibilidade variando de 86.2% a 34.5%, e de especificidade entre 87% e 26.1%. Ao comparar o índice de GM sérica nos animais do grupo caso quanto a presença ou não de anticorpos detectados pela IDGA foi encontrado p=0.503. A detecção de GM pelo teste Platelia Aspergillus EIA® não parece ser útil para o diagnóstico da aspergilose em pinguins naturalmente infectados.

Diagnostic value of the combinations of bronchoalveolar lavage fluid galactomannan test and serum galactomannan test in invasive pulmonary aspergillosis / 中华危重病急救医学

OBJECTIVE@#To evaluate the accuracy and diagnostic value of bronchoalveolar lavage fluid galactomannan test (BALF-GM) combined with serum GM test on invasive pulmonary aspergillosis (IPA).@*METHODS@#190 cases of BALF-GM and 4 787 cases of serum GM specimens suspected of fungal infection in patients admitted to Affiliated Hospital of Jining Medical University from January 2016 to June 2018 were enrolled and analyzed. All patients were classified into clinically confirmed IPA, clinically diagnosed IPA, suspected IPA and excluded IPA according to the classification standard of Expert consensus on diagnosis and treatment of pulmonary mycosis. The coincidence rate of BALF and serum GM test results with clinical diagnosis was analyzed. Receiver operating characteristic (ROC) curve was performed, and the diagnostic value of BALF and serum GM test alone or in combination for IPA was evaluated. Subgroup analysis was performed in patients with normal or abnormal immune function, and the sensitivity and specificity of BALF and serum GM test were compared separately or jointly.@*RESULTS@#The positive rate of BALF-GM was 46.8% (89/190), and 10.4% (497/4 787) on serum GM. Among them, 156 patients were both tested on BALF and serum GM. There were 44 cases with both positive in BALF and serum GM, the coincidence rate of clinical definite was 93.2% (41/44). There were 34 cases with positive BALF-GM and negative GM test in serum, and the coincidence rate of clinical definite was 64.7% (22/34). There were 56 cases positive in serum GM and negative in BALF-GM, and the coincidence rate of clinical definite was 48.2% (27/56). BALF and serum GM tests were both negative in 22 cases, and the coincidence rate of exclusion diagnosis was 90.9% (20/22). ROC curve analysis showed that the diagnostic value of BALF-GM test combined with serum GM test for IPA was better than that of BALF-GM test or serum GM test alone [area under ROC curve (AUC): 0.992 vs. 0.983, 0.976]. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 95.3%, 87.0%, 93.2% and 90.9%, respectively. Subgroup analysis showed that among 89 patients with positive BALF-GM test, 85 cases (95.5%) had normal immune function and 4 cases (4.5%) had unknown condition. Among 497 patients with positive serum GM test, 12 cases (2.4%) had normal immune function, 372 cases (74.9%) had abnormal immune function and 113 cases (22.7%) were uncertain. It was shown by ROC curve analysis that the sensitivity of positive BALF-GM test in diagnosis of IPA in patients with normal immune function was higher than that of positive serum GM test (95.6% vs. 88.9%), while the sensitivity of positive serum GM test in patients with abnormal immune function was higher than that of positive BALF-GM test (91.8% vs. 89.9%).@*CONCLUSIONS@#The results of BALF and serum GM tests are in good agreement with clinical diagnosis, and the combined detection of BALF and serum GM is more valuable for IPA diagnosis than single detection, especially for patients with unknown immune function.

Diagnostic Value of G and GM Tests in Acute Leukemia Patients with Invasive Fungal Disease / 中国实验血液学杂志

OBJECTIVE@#To analyze the diagnostic value of (1, 3) -β-D-glucan and galactomannan (GM) tests in the patients with acute leukemia complicated by invasive fungal disease, and explore the application of serological detection (G/GM) and lung CT for early diagnosis of invasive fungal disease (IFD).@*METHODS@#A total of 493 patients with acute leukemia complicated by high risk invasive fungal infection, also receival G and GM tests, in Department of hematology of our hospital from June 2016 to December 2016 were selected and were divided into IFD-confirmed group (62 cases) including confirmed and clinical diagnesed IFD, and IFD-unconfirmed group (431 cases) including suspected IFD and non-IFD according to the diagnostic criteria of IFD. The results of G and GM tests in patients of 2 groups were analyzed, then the diagnostic efficacy of G and GM done and combination evaluated. In addition, 26 patients whose lung CT negative at hospitalization, moreover, presentation of changes in lung by CT during hospitalization and serological G and GM test positive were selected, and the difference of time between serological that postive and presentation of changes in lung by CT were compared for the estimation of early diagnotic value.@*RESULTS@#The positive rate of (1, 3) -β-D-glucan in IFD-confirmed group and IFD-unconfirmed group was 56.5% and 10.4%, respectively. Meanwhile, that of galactomannan test was 41.9% and 9.0%, respectively. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of (1, 3) -β-D-glucan was 56%, 90%, 44% and 92%, and that of galactomannan was 42%、91%、40% and 93%, respectively. Moreover, the combination of (1, 3) -β-D-glucan and galactomannan could raise the sensitivity to 69% and specificity to 98%. The positive results of serological detection (G/GM) come earlier about five days than CT changes.@*CONCLUSION@#Both (1, 3) -β-D-glucan and galactomannan test have high sensitivity and specificity, and the combination of them can improve the diagnostic efficacy, and make the clinical antifungal therapy more precisely. In the early clinical diagnosis of IFD, the positive results of serological detection coming earlier than lung CT.

Aspergilosis pulmonar invasora en pacientes reumatológicos: presentación de dos casos / Invasive pulmonary aspergillosis in rheumatological patients: presentation of two cases

La aspergilosis pulmonar invasora (API) es una infección causada por hongos del género Aspergillus que afecta principalmente a pacientes inmunocomprometidos y corresponde a la forma más grave de aspergilosis. Se asocia a una alta morbi-mortalidad, siendo fundamental un diagnóstico y tratamiento oportuno. Las manifestaciones clínicas son inespecíficas, por lo que un estudio adecuado es importante para el diagnóstico, principalmente en pacientes con factores de riesgo poco habituales. En la actualidad se han establecido categorías diagnósticas que consideran factores del hospedero, laboratorio micológico tradicional y biomarcadores como galactomanano. Éstos, junto a la mejor comprensión e interpretación de las imágenes tomográficas permiten ofrecer un manejo adecuado. En este artículo, se presentan dos casos clínicos de API en pacientes reumatológicos, y se discute la utilidad de los métodos diagnósticos.

Invasive pulmonary aspergillosis (IPA) is an infection caused by fungi of the genus Aspergillus that mainly affects immunocompromised patients and corresponds to the most severe form of aspergillosis. It is associated with high morbidity and mortality, and diagnosis and timely treatment are essential. Clinical manifestations are nonspecific, so an adequate study is important for diagnosis, mainly in patients with unusual risk factors. At present, diagnostic categories have been established that consider factors of the host, traditional mycological laboratory and biomarkers such as galactomannan. These, together with the better understanding and interpretation of the tomographic images, allow us to offer an adequate management. In this article, two clinical cases of API in rheumatological patients are presented, and the usefulness of the diagnostic methods is discussed.

Can Aspergillus fumigatus conidia cause false-positive results in the galactomannan enzyme immunoassay test?

Abstract INTRODUCTION: Several factors can cause false-positive results in the galactomannan (GM) test; however, others remain unknown. Presently, the impact of airborne contamination by Aspergillus conidia during enzyme-linked immunosorbent assay (ELISA) remains uninvestigated. METHODS: We studied 12 A. fumigatus isolates. Fungal conidia were serially diluted and tested for GM detection using the Platelia® Aspergillus enzyme immunoassay (EIA). RESULTS: The conidia concentration required for an EIA-positive result was 4.8 × 103 (median). CONCLUSIONS: This is the first study to evaluate the impact of environmental contamination on the Platelia® Aspergillus EIA assay. Only massive contamination can interfere with GM optical readings, suggesting that environmental contamination does not cause false-positive test results.

Galactomannan use in clinical practice: providing free testing is not the full answer

ABSTRACT Introduction: Invasive aspergillosis is a condition associated with a high mortality rate mostly due to difficulties in performing an early diagnosis. In recent years, galactomannan detection has markedly improved the diagnosis of invasive aspergillosis, but very little is known on how physicians deal with this test in clinical practice. Methods: This cross-sectional study aimed to analyze the indications for the use of serum galactomannan in a large Brazilian hospital, between 2015 and 2016. No specific protocol was in place for GM request. We reviewed the medical records of adult (>18 years-old) patients who were tested for galactomannan due to one the following indications: screening, diagnosis, or treatment follow-up. Additional variables included demographic data, underlying diseases, presence of neutropenia, and use of previous antifungal (anti-Aspergillus) drugs. Results: The mean age of the patients was 51 years-old (sd ± 15.8), and 63.3% of patients were male. Patients with hematological malignancies accounted for 60.1% of the cases, mostly acute myeloid leukemia (19.6%). Galactomannan testing was positive in 12.2% of patients, including 1.6% of occasions in which the test was used for screening purposes, 13.2% for diagnosis, and 32.4% during follow-up. Median time for chest imaging request was two days before GM testing. Previous antifungal therapy was reported for 35.1% of patients, mostly amphotericin B (57.1%). Conclusion: The correct use of galactomannan testing is essential for an early diagnosis of invasive aspergillosis, which may improve the prognosis of the disease. We demonstrated that clinicians usually ask for galactomannan tests to confirm imaging findings in patients who frequently were on antifungal drugs, something that could be improved by medical education. We observed a low frequency of galactomannan use for preemptive antifungal therapy (25.7%), which is worrying considering the well-known beneficial use of GM testing in this scenario.

Teste de galactomanana para o diagnóstico de aspergilose invasiva: uma revisão / Galactomannan test for the diagnosis of invasive aspergillosis: a review

As doenças fúngicas invasivas têm sido um problema crescente em ambientes hospitalares, sobretudo nas últimas duas décadas. A aspergilose invasiva (AI), ocasionada pelo gênero Aspergillus, está entre as principais causas de morte em pacientes gravemente imunocomprometidos, com mortalidade que varia de 70 a 90%. O padrão de referência para o diagnóstico de AI é o cultivo do micro-organismo e a análise histopatológica dos órgãos afetados. Estes procedimentos são dificilmente realizados na maioria dos casos, e apresentam baixa sensibilidade (<50%), além de as amostras serem habitualmente obtidas em estados avançados da infecção. O teste de detecção de galactomanana tem sido objeto de estudo para o diagnóstico de AI, por representar uma promissora ferramenta e por ser uma técnica sorológica rápida e não invasiva. A presente revisão tem por objetivo fazer levantamento de estudos que utilizaram o teste de galactomanana em amostras de pacientes com quadros clínicos distintos, porém com suspeita e/ou com comprovada AI, bem como as atuais tendências de conhecimento, aplicação e utilidade do ensaio laboratorial.

Invasive fungal diseases represent an increasing problem in the hospital environments, predominantly in the last two decades. The invasive aspergillosis (IA), induced by Aspergillus species, has been the main cause of death in severely immunocompromised patients, with mortality varying from 70 to 90%. Difficulties are found for diagnosing the IA. In vitro culture of biological material shows low sensitivity (<50%), besides the positivity usually occurs at the advanced stages of the infection. The test for detecting galactomannan has been the object of the present study, seeing that it represents a promising diagnostic tool, as a fast and non-invasive serological procedure. The objective of the present review is to survey the studies which have been performed by using methods for detecting galactomannan in samples from patients with distinct clinical pictures. Patients presenting suspicion and/or confirmed IA were also included, as well as the up-to-date trends in knowledge, application and utility of the test.

Depolymerized konjac glucomannan: preparation and application in health care / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Konjac glucomannan (KGM) is a water-soluble polysaccharide obtained from the roots and tubers of konjac plants. Recently, a degraded product of KGM, depolymerized KGM (DKGM), has attracted attention because of its low viscosity, improved hydrophily, and favorable physiological functions. In this review, we describe the preparation of DKGM and its prebiotic effects. Other health benefits of DKGM, covering antioxidant and immune activity, are also discussed, as well as its safety. DKGM could be a candidate for use as a tool for the treatment of various diseases, including intestinal flora imbalance, and oxidative- and immune-related disorders.

Evaluación del Aspergillus lateral flow device para el diagnóstico de aspergilosis invasora, experiencia en un hospital universitario / Evaluation of the lateral flow Aspergillus assay for the diagnosis of invasive aspergillosis, experience in a university hospital

Resumen Introducción: El diagnóstico de aspergilosis invasora (AI) se realiza mediante criterios clínicos y microbiológicos los que incluyen marcadores séricos. Recientemente, el test inmunocromatográfico Aspergillus lateral flow device (LFD), ha sido evaluado como método para diagnóstico de AI. Objetivo: Evaluar el desempeño de este test para el diagnóstico de AI. Material y Método: Estudio transversal en que se evaluaron muestras de suero y lavado bronco-alveolar (LBA) procesadas para galactomanano provenientes de pacientes adultos con sospecha de AI, atendidos en el Hospital Clínico de Red de Salud UCCHRISTUS. Resultados: Se procesó un total de 142 muestras de 98 pacientes, correspondientes a AI probada 5,6%, AI probable 41,5%, AI posible 12,7% y ausencia de AI 40,1%. Al confrontar los resultados con las categorías diagnósticas según criterios EORTC/MSG se obtuvo una sensibilidad y especificidad de LFD para diagnóstico de AI de 70,9 y 53,5% para muestras de suero y 83,3 y 38,5% para muestras de LBA. La concordancia entre galactomanano y LFD fue de 62,4% (54,1-69,9) con un índice Kappa de 0,202 (0,03682-0,3669). Conclusiones: Aspergillus LFD presentó una adecuada sensibilidad; sin embargo, la especificidad fue baja por lo que un resultado positivo requiere ser confirmado.

Background: The incidence of invasive aspergillosis is increasing. Its diagnosis is based on clinical and microbiological criteria which include the determination of serological markers such as galactomannan. Recently, the Aspergillus lateral flow device, an inmunocromatograph assay has been described for its diagnosis. Aim: To evaluate the performance of the lateral flow device for the diagnosis of invasive aspergillosis (IA) in adult patients. Material and Method: In this cross-sectional study, frozen samples that had been previously evaluated for galactomannan from patients classified with proven/probable/possible or no AI according to the EORTC/MSG criteria were selected. Results: A total of 142 samples from 98 patients were processed, corresponding to proven AI 5.6%, probable IA 41.5%, possible IA 12.7% and no-IA 40.1%. The sensitivity and specificity of the Aspergillus lateral flow was 70.9% and 53.5% for serum samples and 83.3% and 38.5% for BAL samples. The concordance between the galactomannan and Aspergillus lateral flow was 62.4% (54.1 - 69.9) with a Kappa index of 0.202 (0.03682 - 0.3669). Conclusions: We observed a good sensitivity but low specificity, a positive result need a confirmatory test.

Serum markers as an aid in the diagnosis of pulmonary fungal infections in AIDS patients

ABSTRACT Introduction: The etiology of pulmonary infections in HIV patients is determined by several variables including geographic region and availability of antiretroviral therapy. Materials and methods: A cross-sectional prospective study was conducted from 2012 to 2016 to evaluate the occurrence of pulmonary fungal infection in HIV-patients hospitalized due to pulmonary infections. Patients' serums were tested for (1-3)-β-D-Glugan, galactomannan, and lactate dehydrogenase. The association among the variables was analyzed by univariate and multivariate regression analysis. Results: 60 patients were included in the study. The patients were classified in three groups: Pneumocystis jirovecii pneumonia (19 patients), community-acquired pneumonia (18 patients), and other infections (23 patients). The overall mortality was 13.3%. The time since diagnosis of HIV infection was shorter in the pneumocystosis group (4.94 years; p = 0.001) than for the other two groups of patients. The multivariate analysis showed that higher (1-3)-β-D-Glucan level (mean: 241 pg/mL) and lactate dehydrogenase (mean: 762 U/L) were associated with the diagnosis of pneumocystosis. Pneumocystosis was the aids-defining illness in 11 out of 16 newly diagnosed HIV-infected patients. Conclusion: In the era of antiretroviral therapy, PJP was still the most prevalent pulmonary infection and (1-3)-β-D-Glucan and lactate dehydrogenase may be suitable markers to help diagnosing pneumocystosis in our HIV population.

Invasive fungal infection in patients with hematologic disorders in a Brazilian tertiary care hospital

ABSTRACT INTRODUCTION: Invasive fungal infections (IFIs) are an important complication in immunocompromised individuals, particularly neutropenic patients with hematological malignancies. In this study, we aimed to verify the epidemiology and diagnosis of IFIs in patients with hematologic problems at a tertiary hospital in Goiânia-GO, Brazil. METHODS: Data from 117 patients, involving 19 cases of IFIs, were collected. The collected data included diagnosis methods, demographics, clinical characteristics, and in vitro susceptibility to different antifungal agents. Among the 19 cases, 12 were classified as proven IFI and 7 as probable invasive aspergillosis with detection of galactomannan in blood and presence of lung infiltrates in radiographic images. Logistic regression analysis showed that the proven and probable IFIs were associated with increased risk of death. Statistical analysis demonstrated that age, sex, and underlying disease were not independently associated with risk of death in IFI patients. RESULTS: Most bloodstream isolates of Candida spp. exhibited low minimum inhibitory concentrations (MICs) to all antifungal agents tested. Voriconazole and amphotericin had the lowest MICs for Aspergillus spp. and Fusarium spp., but Fusarium spp. showed the least susceptibility to all antifungals tested. Amphotericin B, fluconazole, and itraconazole were found to be inactive in vitro against Acremonium kiliense; but this fungus was sensitive to voriconazole. CONCLUSIONS: Considering the high number of IFI cases, with crude mortality rate of 6%, we could conclude that IFIs remain a common infection in patients with hematological malignancies and underdiagnosed ante mortem. Thus, IFIs should be monitored closely.

Diagnostic-driven antifungal therapy in neutropenic patients using the D-index and serial serum galactomannan testing

Abstract Introduction Invasive mold disease is an important complication of patients with hematologic malignancies, and is associated with high mortality. A diagnostic-driven approach has been an alternative to the classical empiric antifungal therapy. In the present study we tested an algorithm that incorporated risk stratification using the D-index, serial serum galactomannan and computed tomographic-scan to guide the decision to start antifungal therapy in neutropenic patients. Patients and methods Between May 2010 and August 2012, patients with acute leukemia in induction remission were prospectively monitored from day 1 of chemotherapy until discharge or death with the D-index and galactomannan. Patients were stratified in low, intermediate and high risk according to the D-index and an extensive workup for invasive mold disease was performed in case of positive galactomannan (≥0.5), persistent fever, or the appearance of clinical manifestations suggestive of invasive mold disease. Results Among 29 patients, 6 (21%), 11 (38%), and 12 (41%) were classified as high, intermediate, and low risk, respectively. Workup for invasive mold disease was undertaken in 67%, 73% and 58% (p = 0.77) of patients in each risk category, respectively, and antifungal therapy was given to 67%, 54.5%, and 17% (p = 0.07). Proven or probable invasive mold disease was diagnosed in 67%, 45.5%, and in none (p = 0.007) of high, intermediate, and low risk patients, respectively. All patients survived. Conclusion A risk stratification using D-index was a useful instrument to be incorporated in invasive mold disease diagnostic approach, resulting in a more comprehensive antifungal treatment strategy, and to guide an earlier start of treatment in afebrile patients under very high risk.

Development of pH sensitive nanoparticles for intestinal drug delivery using chemically modified guar gum co-polymer

The aim of the research work was to chemically modify guar gum[GG] as a pH sensitive co-polymer and formulating intestinal targeting ESO nanoparticles [NPs] using the synthesized co-polymer. Poly acrylamide-grafted-guar gum [PAAm-g-GG] co-polymer was synthesized by free radical polymerization. Chemical modification of PAAm-g-GG by alkaline hydrolysis results in formation of a pH-sensitive co-polymer. The effect of GG and acryl amide [AAm] on grafting was studied. Esomeprazole magnesium [ESO] loaded pH sensitive NPs were prepared by nano-emulsification polymer crosslinking method and characterized. Sixteen formulations were prepared and the concentration of process variables was varied to obtain nanoparticles of 200-600nm. The NPs were found to be homogenous in size distribution. The encapsulation efficiency and drug loading ranged from 33.2% to 50.1% and 12.2% to 17.2% respectively. Particle size, encapsulation efficiency and drug loading increased along with co-polymer concentration. In-vitro release studies at pH 1.2 for 2 h, followed by pH 6.8 showed that environment pH significantly affected the drug release. SEM has shown that NPs are spherical with smooth surface. The pH sensitive PAAm-g-GGNPs resisted the initial release of the drug from the drug loaded NPs in acidic pH and delayed the release process to a longer period in alkaline environment

Clinical analysis of 16 cases of invasive pulmonary aspergillosis in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the diagnosis and treatment of invasive pulmonary aspergillosis (IPA) in children.</p><p><b>METHOD</b>The clinical data of 16 cases of proven or probable IPA who had been in our Hospital from January 2006 to June 2014 were retrospectively analyzed.</p><p><b>RESULT</b>Among the 16 patients, 11 were males and 5 were females. One child had proven IPA and 15 children had probable IPA. Host risk included long duration use of multiple broad-spectrum antibiotics in 16 cases, neutropenia in 9 cases, invasive mechanical ventilation in 3 cases, primary immunodeficiency disease in 2 cases, long-term use of glucocorticoids in 2 cases, measles in 2 cases, and congenital pulmonary hypoplasia in 1 case. Fever, cough and expectoration were present in all the children with IPA. At the time of diagnosis, the halo sign and subpleural wedge consolidation shadows were more common in neutropenia group (5/9, 7/9) than those in non-neutropenia group(0/7, 1/7)(P<0.05). The cavities and"air-crescent sign"were more common after 15 days to 1 month when the children had been treated with anti-aspergillosis drugs than that at the onset of diagnosis of IPA (P<0.05). The positive rate of serum galactomannan (GM) test was higher than that of sputum culture and serum G test (P<0.05). Thirteen children received voriconazole, in 7 of the children the treatment was effective.</p><p><b>CONCLUSION</b>Neutropenia were the common host risk factors in children with IPA. Subpleural wedge consolidation shadows, the halo sign and the"air-crescent"sign were highly suggestive of the diagnosis of IPA in children. Subpleural wedge consolidation shadows and the halo sign were more common in neutropenia group than in non-neutropenia group in the early stage of the course. Serum GM test played an important role in the diagnosis of IPA in children. Voriconazole was effective in majority of the children with IPA.</p>

Mananoligossacarídeo e ß-glucano em dietas de leitões desmamados / Mannan Oligosaccharides and ß-glucan in diets for weaned piglets

Avaliou-se o efeito da suplementação de mananoligossacarídeo, ß-glucano e antibiótico em dietas de leitões machos castrados, durante a fase de creche (21 aos 54 dias de idade), sobre o desempenho, características morfo-histológicas da mucosa intestinal e ocorrência de diarreia. Foram utilizados 368 leitões de mesma linhagem, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos (1: 330g de mananoligossacarídeo t/ração (oriundos do núcleo), 2: 1.830g de mananoligossacarídeo t/ração (330g oriundos do núcleo + 1.550g da suplementação), 3: 330g de mananoligossacarídeo (oriundos do núcleo) + 500g de ß-glucano t/ração e 4: 330g de mananoligossacarídeo (oriundos do núcleo) + 250g de Colistina t/ração) e quatro repetições com 23 animais por unidade experimental. O desempenho foi avaliado pelo ganho de peso, consumo de ração e conversão alimentar. As características morfo-histológicas da mucosa intestinal estudadas foram altura de vilosidade, profundidade das criptas intestinais, perímetro de vilosidade e a relação altura de vilosidade:profundidade de cripta do duodeno, jejuno e íleo. A ocorrência de diarreia foi observada diariamente pela avaliação de escore fecal. A suplementação de mananoligossacarídeo, ß-glucano e antibiótico na dieta de leitões machos castrados, na fase de creche, não influenciaram o desempenho e a ocorrência de diarreia. Maior altura de vilosidade e maior profundidade de criptas no duodeno e íleo foram verificadas nos animais suplementados com ß-glucano. No jejuno foi observado maior perímetro de vilosidade nos animais suplementados com ß-glucano e nos animais que não receberam suplemento adicional na dieta.(AU)

The aim of this study was to evaluate the effect of supplementation with mannan oligosaccharides, ß-glucan and antibiotic in diets for castrated male piglets during the nursery phase (21 to 54 days of age) on performance, morpho-histological characteristics of the intestinal mucosa and occurrence of diarrhea. A total of 368 piglets of the same strain, distributed in a completely randomized design with four treatments (1- 330 g mannan oligosaccharides/t of diet (derived of nucleus); 2 - 1,830 g mannan oligosaccharides/t of diet (330g derived of nucleus + 1550g of supplementation); 3 - 330g mannan oligosaccharides (derived of nucleus) + 500g ß-glucan/t of diet; and 4 - 330g mannan oligosaccharides (derived of nucleus) + 250g colistin/t of diet) and four replications with 23 animals per experimental unit. Performance was assessed through weight gain, feed intake and feed conversion. The morpho-histological characteristics of the intestinal mucosa studied were villus height, intestinal crypt depth, villus circumference and villus height:crypt depth of duodenum, jejunum and ileum ratio. The occurrence of diarrhea was observed daily by the evaluation of fecal score. Supplementation of mannan oligosaccharides, ß-glucan and antibiotic in the diet of castrated male piglets in the nursery phase did not affect performance and occurrence of diarrhea. Greater villus height and crypt depth in the duodenum and ileum were verified in the animals supplemented with ß-glucan. In the jejunum, greater villus circumference was observed in the animals supplemented with ß-glucan and in the animals that did not receive additional supplemented diet.(AU)

Utilidad de la prueba Aspergillus-LFD para el diagnóstico de aspergilosis: primera experiencia en Chile / Utility of Aspergillus-LFD: first experience in Chile

Introduction: Invasive fungal diseases (IFD) by filamentous fungi are a common cause of morbidity and mortality in immunocompromised patients, especially those with myeloid leukemia. In 2011 a protocol for the rapid diagnosis of IFD by filamentous fungi was implemented in Valparaiso Region. Objectives: To describe cases of IFD by filamentous fungi of the Valparaíso Region, since the implementation of rapid diagnosis and to compare results with the period 2004-2009. Materials and Method: Descriptive and prospective study conducted in two public hospitals: Carlos van Buren at Valparaiso and Gustavo Fricke at Viña del Mar. We selected patients with a diagnosis of filamentous fungal diseases considering the EORTC/MSG criteria. Demographics, underlying diseases, risk factors for EFI, galactomannan (GM) results in blood and bronchoalveolar lavage, cultures and biopsies, treatment and overall lethality rates at 30 days were registered. Results: Eighteen patients were detected, 6 with proven and 12 probable IFD. Nine were diagnosed by GM, 8 by culture and two with both methods. In cases which the agent (9/18) was isolated from Rhizopus oryzae was the most frequent. When comparing overall lethality with the period 2004-2009, there was a reduction of 47.8%, which was statistically significant. Conclusions: Compared to data previously published in the region, demographic and comorbidities of patients with IFD caused by filamentous fungi are similar, however the currently rapid diagnosis protocol has improved survival of patients and lethality experienced overall decrease.

Introducción: la enfermedad fúngica invasora (EFI) por hongos filamentosos es una causa frecuente de morbilidad y mortalidad en pacientes inmunocomprometidos, en especial en aquellos con leucemia mieloide. En el 2011 se implementó en la Región de Valparaíso un protocolo de diagnóstico rápido de la EFI por hongos filamentosos. Objetivos: describir los casos de EFI por hongos filamentosos de la Región de Valparaíso, desde la implementación del diagnóstico rápido y compararlos con el período 2004-2009. Materiales y Método: Estudio descriptivo y prospectivo realizado en los hospitales públicos Carlos van Buren de Valparaíso y Gustavo Fricke de Viña del Mar. Se seleccionaron aquellos pacientes con diagnóstico de EFI por hongos filamentosos considerando los criterios EORTC/MSG. Se obtuvieron datos demográficos, enfermedad de base, factores de riesgo para EFI, resultados de galactomanano (GM), cultivos y biopsias, tratamiento y letalidad global a 30 días. Resultados: Se identificaron 18 pacientes, seis con EFI probadas y 12 probables. Nueve fueron diagnosticados con galactomanano, ocho con cultivos y uno con los dos métodos. En los casos en que se aisló el agente (9/18), Rhizopus oryzae fue el más frecuente. Al comparar la letalidad global con la del período 2004-2009, hubo una reducción de 47,8%, la cual fue estadísticamente significativa. Conclusiones: En relación a lo publicado anteriormente en la región, se conservan las características demográficas y de co-morbilidad de los pacientes con EFI por hongos filamentosos; sin embargo, la introducción del nuevo protocolo de diagnóstico rápido se asoció a una disminución en la letalidad global.

Early serological diagnosis of invasive aspergillosis in renal transplant recipients / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the clinical value of serum 1,3-beta-D-glucan (BG) and galactomannan (GM) detection for early diagnosis of invasive aspergillosis (IA) in patients after renal transplantation.</p><p><b>METHODS</b>Blood samples collected from 69 renal transplant recipients were divided into diagnosis group, clinical diagnosis group, suspected diagnosis group, and non-infected group for detection of serum BG and GM.</p><p><b>RESULTS</b>The mean serum levels of BG in the diagnosis group, clinical diagnosis group, and suspected diagnosis group were significantly higher than that in non-infected group (P<0.05). The sensitivity, specificity, and positive and negative predictive values of BG was 69.49%, 70%, 93.18% and 35.71% for IA diagnosis, respectively. The serum levels of GM in the 3 diagnosis groups were also significantly higher than that in the non-infected group (P<0.05) with the sensitivity, specificity, and positive and negative predictive values of 84.75%, 90%, 96.15% and 52.63% for IA diagnosis, respectively.</p><p><b>CONCLUSION</b>Increased serum BG and GM levels can serve as the evidence for early diagnosis of IA with a high diagnostic sensitivity and specificity in renal transplant recipients.</p>