The serodiagnosis of hydatid disease is a valuable instrument for clinical diagnosis and epidemiological surveillance of high-risk populations. In the past decade a wealth of reports on the diagnostic performance of numerous antigens have been produced. However, their diagnostic value has been estimated under different conditions, using different serum collection, therefore precluding their direct comparison. Here we report an unbiased comparison of the same batch of six major E. granulosus antigens, namely, hydatid cyst fluid (HCF), native antigen B (AgB), two recombinant AgB subunits, an AgB-derived synthetic peptide, and recombinant cytosolic malate dehydrogenase from E. granulosus (EgMDH), against the same serum collection. The doubleblind analysis was performed using a standardized protocol and receiver operating characteristic (ROC) data analysis by a network of six South American laboratories. High intercenter reproducibility was attained, and the intralaboratory analysis allowed the comparative ranking of the antigen panel. HCF, AgB, and its AgB8/1 subunit exhibited equivalent diagnostic efficiencies, 81.4% ؎ 0.5%, 81.3% ؎ 0.6%, and 81.9% ؎ 2.0%, respectively; with a more favorable balance toward specificity in the case of the last antigen. The diagnostic efficiencies for the other three antigens were 76.8% ؎ 6.8%, 69.1% ؎ 2.7%, and 66.8% ؎ 2.1%, for the peptide, the AgB8/2 subunit, and the EgMDH, respectively. The study also included an analysis of batch-to-batch variation in the diagnostic performance of different HCF regional preparations. Based on these results, a suggested recommendation on the use of these antigens was drawn.

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SUMMARYFascioliasis is an emerging/re-emerging vector-borne disease with the widest known distribution. Approximately 17 million people are infected around the world, being the Andean region the most affected area. There is an important necessity to develop sensitive and specific diagnostic tools to treat patients early and to avoid complications. In this paper we evaluated the immune response of infected humans against two antigenic preparations: the total soluble extract (FhTSE) and the adult worm vomit (FhAWV) in order to identify antigenic fractions specific for Fasciola hepatica. Both preparations were processed by SDS-PAGE and Western blot with human sera with fascioliasis (F), other parasitosis and healthy individuals. In the immunoblot of FhTSE, sera F recognised 16 bands with MW between eight and 110 kDa, from which those of 8, 9, 10, 38, 45 and 57 kDa were specific. In the preparation FhAWV, sera F recognised nine bands with MW from eight to 85 kDa, from which those of 8, 12, 15 and 24 kDa were specific. Some bands of cross-reaction were evident with sera from patients with other parasitoses, more frequent with the FhTSE. Bands within the MW mentioned, particularly that of eight kDa, have been shown to be specific by others, and deserve additional characterisation for their potential use in immunodiagnosis.

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Purpose Rhesus-(Rh-) negative women receiving anti-D antibodies antenatally often have a positive antibody screen at delivery. We investigated the incidence of positive antibody screens at delivery in this population and examined how the presence of positive antibody screens affected the time required to obtain type and screen or type and crossmatch results. Methods Records of parturients who had type and screen or type and crossmatch done upon presentation for delivery from June to October 2007 were examined to determine estimated gestational age at admission, Rh-status, the presence of positive antibody screens, and the time interval from receipt of specimen in the blood bank to the availability of antibody screen results. Results Of the 480 specimens sent for type and screen or type and crossmatch, 20% of parturients were Rh-negative, with 57% of those demonstrating a positive antibody screen compared with 4% of the Rh-positive parturients (P \ 0.01). In the Rh-negative group, 100% (95% CI 98-102) of positive antibody screens were anti-D antibodies. There was a longer median laboratory time for Rh-negative vs Rh-positive parturients (146 vs 65 min), for antibody positive vs antibody negative parturients (243 vs 65 min) (P \ 0.001 for both), but not for Rh-positive/antibody positive vs Rh-negative/antibody positive patients (312 vs 218 min) (P = 0.09). The antibody screen was positive in 100% of Rh-negative parturients until 37 weeks gestation, after which there was a decline. Conclusions Rh-negative parturients who receive anti-D antibodies antenatally have a higher incidence of positive antibody screens at delivery than Rh-positive parturients due to the presence of anti-D antibodies.

Citar como: Antitupa I, Quispe W, Mayo J, Valverde F, Sanchez E. Purificación de la fracción antigénica 27-28 KDa a partir del antígeno metabólico secretadoexcretado de Fasciola hepatica. Rev Peru Med Exp Salud Publica. 2014;31(2) RESUMENEn el presente estudio, las fracciones antigénicas de 27-28 KDa de Fasciola hepatica fueron purificadas por cromatografía de exclusión molecular para su aplicación en el diagnóstico de la fascioliasis humana. Se obtuvieron antígenos de excreción y secreción a partir de fasciolas adultas vivas obtenida de hígado de ovino y bovino, y cultivados en medio mínimo esencial. La reactividad y eficacia del antígeno purificado fueron evaluadas por la prueba de inmunoblot empleando cuatro sueros con fascioliasis humana; cuatro sueros con otras parasitosis, y dos sueros negativos. Se concluye que las fracciones antigénicas purificadas no presentan reacción cruzada con otras parasitosis, por inmunoblot, por lo que se considera a las proteínas purificadas como potenciales candidatas a ser utilizadas para el diagnóstico de fascioliasis humana.Palabras clave: Fasciola hepatica; Purificación; Cromatografía (fuente: DeCS BIREME). PURIFICATION OF ANTIGENIC FRACTION 27-28 kDa from the metabolic ANTIGEN FROM METABOLIC secreted-excreted from Fasciola hepatica ABSTRACTAntigenic fractions of 27-28 kDa from Fasciola hepatica were purified by size-exclusion chromatography for use in the diagnosis of human fasciolosis. Excretion and secretion antigens were obtained from living adult flukes collected from sheep and cattle liver, and cultured in minimum essential medium. The reactivity of the purified antigen and efficacy were assessed by immunoblot test using four sera with human fascioliasis; four sera with other parasites, and two negative sera. We conclude that the purified antigenic fractions do not cross-react with other parasites by immunoblot. Therefore, purified proteins are considered as potential candidates to be used for the diagnosis of human fascioliasis.