Multiple-bead assay for the differential serodiagnosis of neglected human cestodiases: Neurocysticercosis and cystic echinococcosis.

BACKGROUND: Neurocysticercosis (NCC), and cystic echinococcosis (CE) are two neglected diseases caused by cestodes, co-endemic in many areas of the world. Imaging studies and serological tests are used in the diagnosis of both parasitic diseases, but cross-reactions may confound the results of the latter. The novel multiplex bead-based assay with recombinant antigens has been reported to increases the diagnostic accuracy of serological techniques. METHODOLOGY: We set-up an immunoassay based on the multiplex bead-based platform (MBA), using the rT24H (against Cysticercus cellulosae, causing cysticercosis) and r2B2t (against Echinococcus granulosus sensu lato, causing CE) recombinant antigens, for simultaneous and differential diagnosis of these infections. The antigens were tested on 356 sera from 151 patients with CE, 126 patients with NCC, and 79 individuals negative for both diseases. Specificity was calculated including sera from healthy donors, other neurological diseases and the respective NCC or CE sera counterpart. The diagnostic accuracy of this assay was compared with two commercial ELISA tests, Novalisa and Ridascreen, widely used in the routine diagnosis of cysticercosis and CE, respectively. MAIN FINDINGS: For the diagnosis of NCC, sensitivity ranged from 57.94-63.49% for the rT24H-MBA, and 40.48-46.03% for Novalisa ELISA depending on exclusion or inclusion of sera having equivocal results on ELISA from the analysis; specificities ranged from 90.87-91.30% and 70.43-76.96%, respectively. AUC values of the ROC curve were 0.783 (rT24H) and 0.619 (Novalisa) (p-value < 0.001). For the diagnosis of CE, the sensitivity of the r2B2t-MBA ranged from 68.87-69.77% and of Ridascreen ELISA from 50.00-57.62%; specificities from 92.47-92.68% and from 74.15-80.98%, respectively. AUC values were 0.717 and 0.760, respectively. CONCLUSIONS/SIGNIFICANCE: Overall, the recombinant antigens tested with the bead-based technology showed better diagnostic accuracy than the commercial assays, particularly for the diagnosis of NCC. The possibility of testing the same serum sample simultaneously for the presence of antibodies against both antigens is an added value particularly in seroprevalence studies for cysticercosis linked to control programs in endemic areas where these two parasites coexist.

Evaluation of the sensitivity and specificity of GST-tagged recombinant antigens 2B2t, Ag5t and DIPOL in ELISA for the diagnosis and follow up of patients with cystic echinococcosis.

Cystic echinococcosis (CE) is a neglected zoonotic disease caused by Echinococcus granulosus sensu lato. Diagnosis and monitoring of CE rely primarily on imaging while serology is used as a confirmatory test. However, imaging is not always conclusive and currently available serological assays have suboptimal sensitivity and specificity, lack standardization, and are not useful for patients´ follow-up. Seroassays for CE are usually based on hydatid fluid (HF), a complex, variable antigenic mixture, and cross-reactivity exists especially with alveolar echinococcosis. Recombinant proteins based on immunogenic antigens most abundant in HF, such as AgB1, AgB2 and Ag5, have been used to overcome these limitations. None of them so far showed potential to replace HF; however, their performance have been largely tested on a limited number of samples, and comparison of different antigens using the same cohort has been rarely performed. The combination of several immunogenic epitopes in a single recombinant protein could enhance test sensitivity. For the diagnosis and follow-up of patients with CE, we compared the performance of the crude HF, previously described recombinant 2B2t antigen, and GST-tagged version of 2B2t, and novel designed recombinants (GST-Ag5t and the GST-DIPOL chimera containing AgB1, AgBB2 and Ag5 epitopes) by IgG-ELISA format. Samples belong to a retrospective cohort of 253 well-characterized patients with CE, previously described for the evaluation of the 2B2t antigen, 92 patients with alveolar echinococcosis, and 82 healthy donors. The reference standard for CE diagnosis was the presence of a CE lesion as diagnosed by ultrasonography. The highest sensitivity was obtained with HF [86.7%, 95% confidence interval (CI): 81.2-91.0], followed by GST-2B2t (70.0%, 95% CI: 63.1-76.2), 2B2t (65.5%, 95% CI: 58.5-72.0), GST-Ag5t (64.5%, 95% CI: 57.5-71.1) and GST-DIPOL (63.1%, 95% CI: 56.0-69.7). The GST-2B2t had the best specificity (95.8%, 95% CI: 88.3-99.1) and the lowest cross-reactivity (38.7%, 95% CI: 27.6-50.6). Good response to treatment also correlated to negative test results in the GST-2B2t ELISA. While none of the tested recombinant antigen appears suitable to replace HF for the diagnosis of CE, GST-2B2t should be further explored as a confirmation test, based on its high specificity and low cross-reactivity, and for the follow-up after treatment in those patients with positive serology for this antigen.

Evaluation of the recombinant antigens B2t and 2B2t, compared with hydatid fluid, in IgG-ELISA and immunostrips for the diagnosis and follow up of CE patients.

Cystic echinococcosis (CE) is one of the most widespread helminthic zoonoses and is caused by the tapeworm Echinococcus granulosus complex. CE diagnosis and monitoring primarily rely on imaging techniques, complemented by serology. This is usually approached by the detection of IgG antibodies against hydatid fluid (HF), but the use of this heterogeneous antigenic mixture results in a variable percentage of false positive and negative results, and has shown to be useless for follow-up due to the long persistence of anti-HF antibodies in cured patients. To improve test performances and standardization, a number of recombinant antigens mainly derived from HF have been described, among them the B2t and 2B2t antigens. The performance of these antigens in the diagnosis and follow up of patients with CE has been so far evaluated on a limited number of samples. Here, we evaluated the performances of tests based on B2t and 2B2t recombinant antigens compared to HF in IgG-ELISA and immunochromatography (IC) for the diagnosis and follow-up of patients with CE in a retrospective cohort study. A total of 721 serum samples were collected: 587 from 253 patients with CE diagnosed by ultrasonography (US), 42 from patients with alveolar echinococcosis and 92 from healthy donors from Salamanca (Spain). The highest overall sensitivity was obtained with HF in ELISA (85.5%), followed by IC containing HF and 2B2t-HF (83.0% and 78.2%, respectively). The lowest sensitivity was obtained with B2t and 2B2t in ELISA (51.8%). The highest specificity was obtained with IC containing 2B2t-HF (100%), and the lowest with HF-ELISA (78.0%). The lowest cross-reactivity with sera from patients with alveolar echinococcosis was detected with the recombinant antigens in ELISA (9.5% - 16.7%) and the highest with the HF-IC (64.3%). The results of B2t and 2B2t-ELISA were influenced by cyst stage, as classified by US according to the WHO-Informal Working Group on Echinococcosis (WHO-IWGE), with low sensitivity for inactive (CE4 and CE5) cysts, and by the drug treatment, with higher sensitivity in patients after drug treatment compared with patients not subjected to drug treatment. The two recombinant antigens in ELISA provided promising results for monitoring patients in follow-up, although their use is limited to patients with positive serology against them at the beginning of the follow-up. Potential biological reasons behind the low sensitivity of the recombinant antigens and possible strategies to enhance the performance of CE serology are discussed.

Uso de macro dosis de oxitocina en mujeres conóbito fetal / Use of macro doses of oxytocin in women with stillbirth / Uso de macro doses de oxitocina em mulheres com óbito fetal

Resumen El objetivo de este estudio fue analizar la mejor evidencia científica disponible sobre el uso de macro dosis de oxitocina comparado con dosis estándar en mujeres que presentaron óbito fetal en el año 2017. Las acciones profesionales obstétricas para la atención en los partos varían ampliamente entre los diferentes países, entre las diferentes instituciones nacionales y los diferentes equipos encargados de dicha atención. El realizar investigación en el campo de obstetricia nos permite determinar cuáles son las mejores acciones basadas en la evidencia científica y de esta manera ofrecer una atención de calidad a la usuaria gestante. Para realizar el estudio se aplicó la metodología de Práctica Clínica Basada en la Evidencia; se inició con la elaboración de la pregunta clínica seguido por búsqueda de información en las bases de datos científicas como MEDLINE, COCHRANE, SCIENCIE DIRECT, TRIPDATA BASE, SCIELO y PUBMED. La búsqueda en todas las bases de datos recuperó un total de 2548 artículos, los cuales fueron revisados cuidadosamente por título y abstract. Al finalizar, se recuperó un artículo el cual fue objeto de análisis crítico. Se concluye que la mejor evidencia disponible es insuficiente para dar respuesta fiable a la pregunta clínica establecida sobre el tema.

Abstract The objective of this study was to analyze the best available scientific evidence on the use of macro doses of oxytocin compared to standard doses in women who presented fetal death in the year 2017. The professional obstetric actions for delivery care vary widely among the different countries, between the different national institutions and the different teams responsible for such care. Conducting research in the field of obstetrics allows us to determine what are the best actions based on scientific evidence and thus provide quality care to the pregnant user. In order to carry out the study, the methodology of Clinical Practice Based on Evidence was applied; It began with the elaboration of the clinical question followed by the search of information in the scientific databases such as MEDLINE, COCHRANE, SCIENCIE DIRECT, TRIPDATA BASE, SCIELO and PUBMED. The search in all the databases retrieved a total of 2 548 articles, which were carefully reviewed by title and abstract. At the end, an article was retrieved which was the subject of critical analysis. It is concluded that the best available evidence is insufficient to give a reliable answer to the clinical question established on the subject.

Resumo O objetivo deste estudo foi analisar as melhores evidências científicas disponíveis sobre o uso de macro doses de oxitocina em comparação com doses padrão em mulheres que apresentaram morte fetal no ano 2017. As ações obstétricas profissionais para atendimento de parto variam amplamente entre as diferentes países, entre as diferentes instituições nacionais e as diferentes equipes responsáveis ​​por esses cuidados. A realização de pesquisas no campo da obstetrícia nos permite determinar quais são as melhores ações baseadas em evidências científicas e, portanto, fornecer cuidados de qualidade para o usuário grávida. Para realizar o estudo, foi aplicada a metodologia da Prática Clínica Baseada na Evidência; Começou com a elaboração da questão clínica seguida pela busca de informações nas bases de dados científicas, como MEDLINE, COCHRANE, SCIENCIE DIRECT, TRIPDATA BASE, SCIELO e PUBMED. A pesquisa em todos os bancos de dados recuperou um total de 2 548 artigos, que foram cuidadosamente revisados ​​por título e resumo. No final, um artigo foi recuperado, que foi objeto de análise crítica. Conclui-se que a melhor evidência disponível é insuficiente para dar uma resposta confiável à questão clínica estabelecida no assunto.

Comparison of T24H-his, GST-T24H and GST-Ts8B2 recombinant antigens in western blot, ELISA and multiplex bead-based assay for diagnosis of neurocysticercosis.

BACKGROUND: Currently, the reference standard assay for the serodiagnosis of neurocysticercosis (NCC) is the lentil lectin-bound glycoproteins/enzyme-linked immunoelectrotransfer blot (LLGP-EITB). The main disadvantage of this technique is the complexity of obtaining and purifying the LLGP extract. This could be solved by replacement with highly specific recombinant antigens from Taenia solium. Based on previous studies, we selected and produced the recombinant Ts8B2 and T24H proteins and applied them to three diagnostic techniques: western blot (WB), enzyme-linked immunosorbent assay (ELISA) and the multiplex bead-based assay (MBA). METHODS: The Ts8B2 and T24H cDNA sequences were expressed in a prokaryotic system and the corresponding expression products purified; three recombinant proteins were further characterized: T24H-his, GST-T24H and GST-Ts8B2. The proteins on WB, ELISA and MBA were tested against 149 sera from patients with NCC confirmed by brain imaging, 40 sera from patients with other parasitic diseases, and 131 sera from US. individuals without evidence of neurocysticercosis (clinical/serological/brain imaging). The sensitivity and specificity of each antigen by WB were calculated by counting the number of true positive, false positive, true negative and false negative results. Using the receiver operating characteristic (ROC) curves, the cut-off values for the ELISA and MBA were established as well as the sensitivity and specificity of each assay. RESULTS: All three antigens showed a high sensitivity on WB in active NCC cases with two or more viable cysts and low sensitivity for cases with single viable cyst or calcified lesions and inactive NCC. WB showed the highest specificity and sensitivity out of the three diagnostic techniques. The recombinant T24H-his was the best diagnostic reagent in WB (100% sensitivity, 99.4% specificity), exhibiting similar results to the LLGP-EITB, against the same panel of NCC sera. The GST-T24H antigen worked better than the others in ELISA and MBA protocols (88.3 and 96.1% sensitivity, respectively and 96.5% specificity). CONCLUSIONS: The sensitivity and specificity that we obtained were similar to results from a previous study using a similar recombinant antigen (rT24H), suggesting that recombinant antigens may be good alternatives to crude extracts in a variety of diagnostic techniques. Furthermore, these antigens can be applied in the development of point-of-care tests which would be useful in NCC field studies.

Follow-up of surgically treated patients with cystic echinococcosis: can novel recombinant antigens compete with imaging? Analysis of a patient cohort.

OBJECTIVE: To compare the performance of two novel recombinant antigens (EgP29, 2B2t) with imaging in a well-defined cohort of surgically treated cystic echinococcosis (CE) patients to determine whether serology reflects surgical cure as defined by imaging. METHODS: From a cohort of 223 CE-confirmed patients of a national clinical center for echinococcosis, 36 surgically treated patients were eligible for analysis. Sera were tested by enzyme-linked immunosorbent assay (ELISA) for specific IgG and IgG4 antibodies against the EgP29 and 2B2t antigens. We used a hierarchical linear regression model to examine the course of antibody levels over time for each patient. A meta-analysis of the patient-specific estimates of the time to negativity was performed using the metan command in Stata. RESULTS: The range of positive serological results at the beginning of post-surgical monitoring was 34-60%: 2B2t 51%, 2B2t-IgG4 34%, EgP29 60% and EgP29-IgG4 40%. The pooled estimates of time to seronegativity were as follows: 2B2t-ELISA 3.92 (3.24, 4.61) years; 2B2t-IgG4-ELISA 4.60 (3.91, 5.29) years; EgP29-ELISA 3.94 (3.50, 4.39) years; EgP29-IgG4-ELISA 2.55 (1.93, 3.18) years. CONCLUSION: After surgical treatment, antibodies to the recombinant antigens 2B2t and EgP29 become negative in the majority of CE-confirmed, surgically cured patients. The major drawback is the fact that only around half of the CE-confirmed, surgically treated patients were at all responsive to the test antigens, so they are of limited benefit for documenting primary cure. Equally, these antigens do not appear to be sensitive to recurrences.

Serological Diagnosis and Follow-Up of Human Cystic Echinococcosis: A New Hope for the Future?

Cystic echinococcosis (CE) is an important helminthic zoonotic disease caused by the Echinococcus granulosus complex. In humans, CE is a chronic disease driven by the growth of echinococcal cysts in different organs. Prognosis of this disease depends on multiple factors, including location, number, size, and stage of the cysts, making CE a disease of complex management. CE is usually asymptomatic for years and attracts limited attention from funding organizations and health authorities. For this reason, only experts' recommendations are available but no evidence-based conclusions have been drawn for CE clinical management. One of those pitfalls refers to the lack of evidence to support the use of serological tools for the diagnosis and follow-up of CE patients. In this respect, crude antigens are used to detect specific antibodies in patients, giving rise to false positive results. The advent of molecular techniques allowing the production of recombinant proteins has provided a number of candidate antigens that could overcome the problems associated with the use of crude parasite extracts in the serological assays. In this review, we present the last advances in this field, proposing the use of serology to support cyst stage-specific diagnosis and follow-up.

A survey of intestinal parasites including associated risk factors in humans in Panama.

BACKGROUND: Intestinal parasitic infections are among the most common infections worldwide, leading to illness with serious and long lasting implications in children and immunocompromised people. Transmission of intestinal parasites is more frequent in tropical and sub-tropical areas where sanitation is poor and socioeconomic conditions are deficient. Panama is a country where climate and social conditions could be reflected in a high number of people infected with intestinal parasites. The presence, prevalence, and distribution of intestinal parasites in this country have been approached to date only in very restricted areas and population groups, but the impact of intestinal parasite infections at the national level is unknown. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a cross-sectional survey between 2008 and 2010 to determine the prevalence of intestinal parasites across Panama. Overall, 14 municipalities in seven provinces of Panama were surveyed. The presence of eggs, cysts, and larvae was assessed by microscopy in 1123 human fecal samples using a concentration technique. A questionnaire to identify risk factors associated with the frequency of intestinal parasites in the study population was also prepared and performed. Overall, 47.4% of human samples presented parasites. Variables including community type, age group, occupation, co-presence of commensals and socioeconomic factors (use of shoes and type of sanitation) were significantly associated with intestinal parasites (p<0.05). CONCLUSIONS/SIGNIFICANCE: The preliminary data obtained in the current study, showing a high prevalence of fecal-oral transmitted parasites in Panama, place intestinal parasitism as a major health problem in this country. Specific interventions should be planned for the indigenous population, the group most afflicted by intestinal parasites.

Improved serodiagnosis of cystic echinococcosis using the new recombinant 2B2t antigen.

A standardized test for the serodiagnosis of human cystic echinococcosis (CE) is still needed, because of the low specificity and sensitivity of the currently available commercial tools and the lack of proper evaluation of the existing recombinant antigens. In a previous work, we defined the new ELISA-B2t diagnostic tool for the detection of specific IgGs in CE patients, which showed high sensitivity and specificity, and was useful in monitoring the clinical evolution of surgically treated CE patients. Nevertheless, this recombinant antigen gave rise to false-negative results in a percentage of CE patients. Therefore, in an attempt to improve its sensitivity, we constructed B2t-derived recombinant antigens with two, four and eight tandem repeat of B2t units, and tested them by ELISA on serum samples of CE patients and patients with related parasites. The best diagnostic values were obtained with the two tandem repeat 2B2t antigen. The influence of several clinical variables on the performance of the tests was also evaluated. Finally, the diagnostic performance of the 2B2t-ELISA was compared with that of an indirect haemagglutination commercial test. The 2B2t recombinant antigen performed better than the HF and B2t antigens, and the IHA commercial kit. Therefore, this new 2B2t-ELISA is a promising candidate test for the serodiagnosis of CE in clinical settings.

Cystic echinococcosis in Spain: current situation and relevance for other endemic areas in Europe.

Cystic echinococcosis (CE) remains an important health problem in many regions of the world, both where no control measures have been implemented, and where control programs have been incompletely successful with ensuing re-emergence of the disease. In Spain, official data on CE show an increase in the proportion of intermediate hosts with CE during the last few years, and autochthonous pediatric patients have been reported, a sign of active local transmission of disease. A similar picture emerges from data reported to the European Food Safety Authority by other European countries. Nevertheless, several crucial aspects related to CE that would help better understand and control the disease have not been tackled appropriately, in particular the emergence of infection in specific geographical areas. In this respect, while some data are missing, other data are conflicting because they come from different databases. We review the current situation of CE in Spain compared with areas in which similar problems in the CE field exist, and offer recommendations on how to overcome those limitations. Specifically, we believe that the introduction of national registries for CE with online data entry, following the example set by the European Registry for Alveolar Echinococcosis, would help streamline data collection on CE by eliminating the need for evaluating and integrating data from multiple regions, by avoiding duplication of data from patients who access several different health facilities over time, and by providing much needed clinical and epidemiological data that are currently accessible only to clinicians.

Usefulness of four different Echinococcus granulosus recombinant antigens for serodiagnosis of unilocular hydatid disease (UHD) and postsurgical follow-up of patients treated for UHD.

Four different recombinant antigens derived from Echinococcus granulosus, designated B1t, B2t, E14t, and C317, were tested with enzyme-linked immunosorbent assays (ELISAs) for the detection of specific immunoglobulin G (IgG) in patients with unilocular hydatid disease (UHD). The results were compared to those obtained with hydatid fluid and were subjected to receiver operator characteristic analysis. The diagnostic performance of the above-listed proteins was defined with respect to their specificity, sensitivity, and predictive values (PV); the influence of cyst location; and usefulness in the follow-up of surgical treatment for UHD and in the determination of whether or not patients have been surgically cured of UHD. The best diagnostic results were obtained with the anti-B2t IgG ELISA, with 91.2% sensitivity, 93% specificity, and high positive and negative PV (89.4 and 94.2, respectively). In addition, this diagnostic tool proved to be useful for the follow-up of surgically treated UHD patients. The anti-B2t IgG ELISA may find an application in the serodiagnosis of UHD in clinical laboratories.

Comunicación entre el personal directivo y operativo de enfermería y su influencia en la productividad / Communication between directing and operating nursing personnel upon their productivity

Se realizó una investigación de campo para saber si la comunicación entre los personales directivo y operativo de enfermería influye sobre la productividad. Se aplicaron dos encuestas, una para cada personal, en el Hospital General de Mexicali de la Secretaría de Salud y en el Hospital de Ginecoobstetricia y Pediatría con Unidad de Medicina Familiar Núm. 31 del Instituto Mexicano del Seguro Social, en la misma ciudad. Los resultados muestran que ambos grupos consideran importante la comunicación para un buen desempeño laboral; sin embargo, reconocen que existen deficiencias en la comunicación, lo que motiva que el personal desconozca los objetivos y programas prioritarios de la institución en que labora. En aspectos como colaboración, satisfacción personal y motivación con el equipo de salud, los resultados fueron positivos, si bien se encontraron incongruencias en lo que se refiere a jornada laboral y carga de trabajo