Assessment of monoclonal antibodies to Echinococcus granulosus Antigen 5 and Antigen B for detection of human hydatid circulating antigens

Parasitology ◽  
1993 ◽  
Vol 106 (1) ◽  
pp. 75-81 ◽  
Author(s):  
D. Liu ◽  
M. D. Rickard ◽  
M. W. Lightowlers
Keyword(s):  
Monoclonal Antibodies ◽  
Echinococcus Granulosus ◽  
Binding Capacity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Antigen B ◽  
Detection Assay ◽  
Circulating Antigens ◽  
Normal Human ◽  
Antigen 5

SUMMARYFour monoclonal antibodies (MAb) to Echinococcus granulosus Antigen 5 (Ag5) and Antigen B (AgB) were assessed in an enzyme-linked immunosorbent assay (ELISA) for detection of circulating antigens (CAg) in sera of human patients with E. granulosus infection. Around 5·5–8% of 200 sera from 42 surgically proven hydatid patients contained detectable CAg by individual MAb. The combined detection rate for CAg, using four MAb, was 19% (38/200). Although hydatid CAg was detected by MAb in at least one serum sample from 21 of 42 patients, some patients remained negative in the assay regardless of the time when serum samples were taken (pre- or post-operatively), or of the continuing presence of hydatid cysts, their location or fertility. In addition, it was observed that the binding capacity of MAb for sheep hydatid cyst fluid antigen (SHCF) was somewhat reduced in the presence of normal human serum. The CAg detection assay would only be useful for assessment of hydatid infection status in patients with detectable CAg in serum samples.

scholarly journals Dynamics of Antigenemia and Coproantigens during a Human Fasciola hepatica Outbreak

1998 ◽  
Vol 36 (9) ◽  
pp. 2723-2726 ◽  
Author(s):  
Ana M. Espino ◽  
Ailén Díaz ◽  
Antonio Pérez ◽  
Carlos M. Finlay
Keyword(s):  
Fasciola Hepatica ◽  
Clinical Symptoms ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
La Palma ◽  
Detection Assay ◽  
Circulating Antigen ◽  
Circulating Antigens ◽  
Stool Samples

In the present study the dynamics of antigenemia and coproantigens were studied in patients withFasciola hepatica infection during an outbreak occurring in La Palma, Pinar del Rı́o, in the West Province of Cuba. Stool and serum samples were collected from 67 patients and 40 healthy subjects. Stool samples were studied by a simple gravity sedimentation technique and an ES78 sandwich enzyme-linked immunosorbent assay (ELISA) for observation of eggs and detection of parasite coproantigens, respectively. Serum samples were also studied by the ES78 sandwich ELISA and an indirect ELISA to detect circulating antigens and antibodies, respectively. At the beginning of the study, 8 of 67 patients had patent infections and 59 had prepatent infections, which was determined by the recent consumption of lettuce contaminated with metacercariae of F. hepatica, the presence of clinical symptoms, and the absence of Fasciolaeggs in their stools. Patients with prepatent infections were monitored by all techniques until patency. Circulating antigens were not detected in patients with patent infections. However, coproantigens were clearly detected in all patients with patent infections. On the other hand, 28.8% of patients with prepatent infections tested positive for circulating antigens and 81.4% tested positive for coproantigens in the first stool sample studied. Only two other coproantigen determinations were necessary to diagnose 93.2% of the patients. While circulating antigen levels diminished in all patients during the infection, coproantigen levels increased. The present study demonstrates that the ES78 sandwich ELISA is a better tool than parasitological examination for diagnosis of active early infection, since by the combination of the circulating-antigen detection assay and the coproantigen detection assay 91% of patients were able to be diagnosed at the beginning of the study. In contrast, a coprologic analysis repeated over several weeks was necessary to diagnose 100% of the patients.

scholarly journals Antibody response in patients with Gaucher disease after repeated infusion with macrophage-targeted glucocerebrosidase

Blood ◽  
1993 ◽  
Vol 82 (5) ◽  
pp. 1402-1409 ◽  
Author(s):  
SM Richards ◽  
TA Olson ◽  
JM McPherson
Keyword(s):  
Gaucher Disease ◽  
Mean Value ◽  
Patient Treatment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Enzyme Replacement ◽  
Human Sera ◽  
Normal Human ◽  
Igg1 Subclass

Abstract Recent clinical data have shown that enzyme replacement therapy with macrophage-targeted glucocerebrosidase (GCR) can be effective in treating type 1 Gaucher disease. Sera from 262 patients, repeatedly infused with GCR, were assessed for the presence of antibodies to this therapeutic protein. Patient serum samples obtained at 3-month intervals were assessed by enzyme-linked immunosorbent assay and those with values greater than two standard deviations above the mean value obtained with a pool of normal human sera were further characterized by radioimmunoprecipitation. At the time of these analyses, the duration of patient treatment varied from 3 months to approximately 3 years. Of the 262 patients analyzed, 34 (12.9%) showed IgG antibodies, as confirmed by radioimmunoprecipitation. All patients who seroconverted did so within 1 year of treatment. The predominant antibody developed was the IgG1 subclass. Fourteen patients in the study experienced periodic symptoms suggestive of immediate hypersensitivity. Nine of these 14 patients had antibody to GCR as determined by radioimmunoprecipitation, whereas 5 patients were antibody negative. There was no evidence of the development of IgE antibodies in these 14 patients. The presence of GCR antibodies did not appear to effect efficacy of therapy in any of the patients treated to date.

Ultrastructural immunocytochemical localization of two hydatid fluid antigens (antigen 5 and antigen B) in the brood capsules and protoscoleces of ovine and equine Echinococcus granulosus and E. multilocularis

Parasitology ◽  
1978 ◽  
Vol 77 (2) ◽  
pp. 143-152 ◽  
Author(s):  
Caroline Davies ◽  
M. D. Rickard ◽  
D. T. Bout ◽  
J. P. Smyth
Keyword(s):  
Echinococcus Granulosus ◽  
Periodic Acid ◽  
Ultrastructural Localization ◽  
Capsule Wall ◽  
Periodic Acid Schiff ◽  
Hydatid Fluid ◽  
Antigen B ◽  
Parenchyma Cells ◽  
Perinuclear Cytoplasm ◽  
Antigen 5

SummaryThe unlabelled antibody method was used in the ultrastructural localization of two hydatid fluid antigens, antigen 5 and antigen B, in brood capsules and protoscoleces of Echinococcus granulosus and E. multilocularis. Antigen 5 was found in the parenchyma cells of the protoscolex and brood capsule wall and to a lesser extent in the walls of the flame cells and collecting ducts of the excretory system and in the surrounding interstitial material. It is suggested that, while some excretion of this antigen may occur from the protoscolex, it could also be liberated into the cystic cavity by degeneration of protoscoleces and parenchymal cells of the brood capsule wall. Antigen B was found mainly in the distal cytoplasm and perinuclear cytoplasm of the tegument anterior to the suckers. It is apparently secreted to the outside and was present in the brood capsule contents; it adheres to the anterior surface and the posterior periodic acid–Schiff (PAS)-positive glycocalyx of the protoscolex and to the inner surface of the brood capsule wall. The protoscolex tegument posterior to the suckers was negative. The parenchyma cells of the protoscolex and brood capsule wall were also positive although the intensity of the reaction product was variable.

scholarly journals Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

2021 ◽  
pp. 2097-2101
Author(s):  
Mohamed J. Saadh ◽  
Samer A. Tanash ◽  
Ammar M. Almaaytah ◽  
Issam J. Sa'adeh ◽  
Saed M. Aldalaen ◽  
...  
Keyword(s):  
Western Blotting ◽  
Clinical Symptoms ◽  
Characteristic Curve ◽  
Fasciola Gigantica ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Cattle ◽  
Serum Samples ◽  
Healthy Cattle ◽  
Target Epitope ◽  
Circulating Antigens

Background and Aim: Diagnosis of fascioliasis depends on clinical symptoms and routine laboratory tests. Recently, antibodies and circulating antigens of Fasciola were used for detecting active infections. Therefore, this study aimed to identify Fasciola gigantica antigens in the sera of infected cattle using Western blotting and enzyme-linked immunosorbent assay (ELISA) for an accurate diagnosis of cattle infected with F. gigantica. Materials and Methods: Serum samples were obtained from 108, 23, and 19 cattle infected with Fasciola gigantica, Paramphistomum cervi, and Strongylids, respectively, including 57 non-infected cattle that were used as healthy cattle for the study. Western blotting and ELISA were then used to detect circulating Fasciola antigens at 27 kDa. Results: The target epitope was detected in an F. gigantica adult-worm antigen preparation, excretory/secretory products, and serum from cattle infected with F. gigantica. However, it was absent in sera from P. cervi, Strongylids, and healthy cattle. The purified 27 kDa F. gigantica (FPA-27) antigen was also detected in cattle serum using ELISA with high degrees of sensitivity and specificity (94% and 82%, respectively), and the area under the receiver operating characteristic curve was 0.89 with a highly significant correlation of p<0.0001. Conclusion: The FPA-27 is proposed to be a promising candidate for the serodiagnosis of fascioliasis in cattle.

scholarly journals Improved Immunodiagnosis of Cystic Hydatid Disease by Using a Synthetic Peptide with Higher Diagnostic Value Than That of Its Parent Protein, Echinococcus granulosus Antigen B

2000 ◽  
Vol 38 (11) ◽  
pp. 3979-3983 ◽  
Author(s):  
Gualberto González-Sapienza ◽  
Carmen Lorenzo ◽  
Alberto Nieto
Keyword(s):  
Hydatid Disease ◽  
Synthetic Peptides ◽  
Diagnostic Value ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Cystic Hydatid Disease ◽  
Antigen B ◽  
Parent Protein ◽  
Diagnostic Sensitivity And Specificity

The assays are used for the diagnosis of hydatid disease are still imperfect. The reported diagnostic sensitivity and specificity vary greatly depending on the panel of sera used, the laboratory conducting the assay, and, more critically, the antigen used. To contribute to its standardization, we have recently ranked the diagnostic performances of the major parasite antigens and the available synthetic peptides using a large collection of serum samples. That work showed that antigen B (AgB) possesses the highest diagnostic value among these antigens. In the present work we further dissected its antigenicity by analyzing the reactivity of the same panel of sera against a set of synthetic peptides spanning the sequence of both AgB subunits. The N-terminal extension of these subunits appeared to be immunodominant in human infections. A 38-mer peptide (p176) delineated from the N-terminal extension of the AgB/1 subunit performed in an enzyme-linked immunosorbent assay with a higher diagnostic sensitivity (80%) and specificity (94%) than native AgB, Ag5, or any other peptide antigen tested against this collection of serum samples. In view of its high diagnostic value and its nature as a well-defined reproducible antigen, p176 could conveniently be used as a reference standard antigen in the diagnosis of hydatid disease.

Erythropoietin Response to Anemia Is Impaired in Patients with Hematologic Malignancies.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3744-3744 ◽  
Author(s):  
Ti Shen ◽  
Yuankai Shi ◽  
Jun Zhu ◽  
Bing Han
Keyword(s):  
Renal Function ◽  
Binding Capacity ◽  
Hematologic Malignancy ◽  
Transferrin Saturation ◽  
Hematologic Malignancies ◽  
Lymphocytic Leukemia ◽  
Deficiency Anemia ◽  
Serum Lactate Dehydrogenase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples

Abstract Anemia is a common complication in patients with hematologic malignancies, either before or after chemotherapy. To find out whether anemic patients with hematologic malignancies have a lower erythropoietin (Epo) response, we analyzed serum Epo levels in these patients. Serum samples for measurement of Epo levels were obtained from 80 patients from three medical centers. Patients had multiple myeloma (MM, n=26), chronic lymphocytic leukemia (CLL, n=6) or non-Hodgkin’s lymphoma (NHL, n=48). Thirty patients had anemia (mean hemoglobin [Hb] level 8.4±1.6 g/dl) and 50 patients were not classed as anemic, with mean Hb levels of 12.7±1.4 g/dl. To provide control values, serum Epo levels were also determined in patients with iron-deficiency anemia (IDA; n=20) and in healthy individuals (n=20). Serum Epo levels were measured by enzyme-linked immunosorbent assay (ELISA). Patients with severe liver or renal dysfunction, low platelet counts or blood loss were excluded. The complete blood cell count (CBC), reticulocyte count, serum lactate dehydrogenase (LDH), liver and renal function, serum iron (SI), total iron-binding capacity (TIBC), transferrin saturation (TSAT), and serum ferritin (SF) levels were evaluated concomitantly. There were no significant differences in CBC, LDH, liver and renal function, SI, TIBC, TSAT and SF values between malignant patients with or without anemia (P>0.05). In patients with hematologic malignancy, those with anemia had higher Epo levels (mean 97.8±183.9 mIU/ml) than those without anaemia (mean 27.8±85.4 mIU/ml; P<0.001). In patients with IDA, the Epo response was inversely correlated with Hb level (r= −0.5, P<0.05), whereas the expected inverse linear relationship between serum levels of Epo and Hb was absent in the group with hematologic malignancies (r= −0.14). Anemic patients with malignancy, and IDA patients both had elevated Epo levels compared with normal controls (means, 97.8±183.9 mIU/ml, 158.3±308.6 mIU/ml and 9.2±4.0 mIU/ml, respectively; both P<0.001 vs controls). However, after correcting for Hb level, anemic patients with malignancy were found to have significantly lower Epo levels than patients with IDA (P=0.032) indicating a decreased Epo response in the former group. These findings indicate that anemia associated with hematologic malignancy may result from an inappropriately low Epo response. Epo treatment should benefit this group of patients.

An evaluation of antigen capture assays for detecting active filarial antigens

2014 ◽  
Vol 89 (3) ◽  
pp. 352-358 ◽  
Author(s):  
R. Ravishankaran ◽  
N.S. Radhika ◽  
L. Ansel Vishal ◽  
S. Meenakshisundaram ◽  
A.A. Karande ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Capture Assay ◽  
Antigen Capture ◽  
Circulating Filarial Antigens ◽  
Detection Assay ◽  
Normal Individuals ◽  
Circulating Antigen ◽  
Endemic Normal

AbstractLymphatic filariasis is a parasitic disease of tropical countries. This is a disfiguring and painful disease contracted in childhood, but the symptoms become apparent only in later years. Diagnosis of filarial infection is very crucial for the management of the disease. The main objective of this study was to develop a filarial antigen-based immunological assay for the diagnosis and surveillance of the disease. Monoclonal and polyclonal antibodies were raised to the recombinant protein Brugia malayi vespid allergen homologue (VAH). Capture enzyme-linked immunosorbent assay (ELISA) was standardized utilizing various combinations of antibodies and evaluated with serum samples of endemic normal (EN, n= 110), microfilaraemic (MF, n= 65), chronic pathology (CP, n= 45) and non-endemic normal (NEN, n= 10) individuals. Of the 230 samples tested, VAH capture assay detected circulating antigen in 97.91% of bancroftian and 100% of brugian microfilaraemic individuals, and 5% of endemic normal individuals, comparable to the earlier reported SXP-1 antigen detection assay. However, the combination of VAH and SXP-1 (VS) capture ELISA was found to be more robust, detecting 100% of microfilaraemic individuals and with higher binding values. Thus an antigen capture immunoassay has been developed, which can differentiate active infection from chronic infection by detecting circulating filarial antigens in clinical groups of endemic areas.

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