Diagnostic Potential of Anti-Rte30 Polyclonal Antibodies in a Blocking Elisa for Toxocara canis Detection

10.1645/17-59 ◽  
2019 ◽  
Vol 105 (1) ◽  
pp. 64
Author(s):  
Cristiane Pires Dias Felicetti ◽  
Francine Sinnott ◽  
Leonardo Garcia Monte ◽  
Karen Leal ◽  
Fabrício Rochedo Conceição ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Toxocara Canis ◽  
Blocking Elisa ◽  
Diagnostic Potential

Comparison of isolates and species ofToxocaraandToxascarisby biosynthetic labelling of somatic and ES proteins from infective larvae

Parasitology ◽  
1991 ◽  
Vol 103 (3) ◽  
pp. 451-464 ◽  
Author(s):  
A. P. Page ◽  
D. T. Richards ◽  
J. W. Lewis ◽  
H. M. Omar ◽  
R. M. Maizels
Keyword(s):  
Polyclonal Antibodies ◽  
Toxocara Canis ◽  
Physicochemical Characteristics ◽  
Toxocara Cati ◽  
Toxocara Vitulorum ◽  
Geographical Regions ◽  
Specific Reactivity ◽  
Species Specific ◽  
Dimensional Electrophoresis ◽  
Toxascaris Leonina

Infective-stage larvae of three different isolates ofToxocara caniswere intrinsically ([35S]methionine) labelled in culture, to determine the presence of similarities or differences in the somatic and ES antigens expressed between larvae derived from different hosts and different geographical regions. Two other closely related ascaridids,Toxascaris leoninawhich infects cats and dogs, andToxocara vitulorum(Neoascaris vitulorum) which infects cattle, were also compared toT. canislarvae by this method. Overall comparisons were made by 1- and 2-dimensional electrophoresis, while immunological cross-reactivities between the different species were analysed by radio-immunoprecipitation. Our results show that extensive physicochemical characteristics are shared betweenT. canisisolates, both from different hosts and different geographical locations. A substantial overlap was revealed whenT. canisandT. vitulorumantigens were compared, whereasToxascariswas found to produce a distinct antigen profile: this result was independent of whether methionine- or Iodogen-labelled products were being considered. Antigen recognition by polyclonal antibodies raised to all three species and to the cat ascarididToxocara cati, revealed considerable cross-reactivities. The cross-reactions were especially prominent between theToxocaraspecies, a fact further substantiated when reactivity ofT. canisES-specific monoclonal antibodies were tested againstT. leoninaandT. vitulorumantigens. The ES antigens ofT. leoninawere not recognized by theT. canismonoclonals, whereas the majority of these antibodies precipitated antigens ofT. vitulorum. One which did not react withT. vitulorumwas monoclonal antibody Tcn 2, indicating its species-specific reactivity and therefore its potential for the specific diagnosis of human toxocariasis.

scholarly journals Immunochemical Characterization of the MPB70/80 and MPB83 Proteins of Mycobacterium bovis

1998 ◽  
Vol 66 (4) ◽  
pp. 1445-1452 ◽  
Author(s):  
Harald G. Wiker ◽  
Konstantin P. Lyashchenko ◽  
A. Murat Aksoy ◽  
Kenneth A. Lightbody ◽  
John M. Pollock ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Homologous Region ◽  
Bacterial Surface ◽  
Purified Protein Derivative ◽  
Diagnostic Potential ◽  
Ring Structures ◽  
Linear Epitopes

ABSTRACT MPB70 and MPB80 (MPB70/80) and MPB83 are closely related antigens which are highly expressed in Mycobacterium bovis. MPB70/80 are soluble secreted antigens, while MPB83 is an exported lipoprotein associated with the bacterial surface. In the present study, these antigens had different mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. These differences may be explained by the fact that MPB70 and MPB83 both have two internal cysteine residues which would create ring structures by disulfide bonding. We analyzed the structures of MPB70/80 and MPB83 by using monoclonal antibodies (MAbs) raised against bovine purified protein derivative or whole M. bovis cells. MAb 1-5C reacted specifically with MPB70 and MPB80, and MAb MBS43 reacted specifically with MPB83, while the other antibodies, including several previously described MAbs, bound all three antigens. MAbs and polyclonal antibodies reacted strongly with reduced protein and less well with nonreduced protein, indicating involvement of linear epitopes. Epitopes of MAbs Bov-1, 2-6B, 1-5C, and 1-1D were mapped by using synthetic peptides of MPB70. Sequence comparison showed the peptide with the 1-5C-reactive epitope to have three residues different from those in the homologous region of MPB83. Exchanges of A for S in position 112 or Q for E in position 116 abolished the reactivity of MAb 1-5C. Polyclonal rabbit antibodies to native purified MPB70 reacted strongly with peptides 6, 7, and 8 of the N-terminal half of mature MPB70. Cattle sera of experimentally M. bovis-infected animals recognized a broader spectrum of peptides. These findings indicate that there is diagnostic potential for these proteins and that there is also a possible role for antibodies in elucidation of the host-mycobacterium relationship involving a surface-bound and exposed lipoprotein, MPB83, and its highly homologous soluble secreted MPB70/80 counterparts.

scholarly journals Diagnostic Potential of Anti-rNcp-43 Polyclonal Antibodies for the Detection of Neospora caninum

2013 ◽  
Vol 68 (4) ◽  
pp. 472-476 ◽  
Author(s):  
Gizele Lima de Sá ◽  
Diene de Borba Pacheco ◽  
Leonardo Garcia Monte ◽  
Francine Alves Sinnott ◽  
Marina Amaral Xavier ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Polyclonal Antibodies ◽  
Diagnostic Potential

scholarly journals Diagnostic Potential of Polyclonal Antibodies Against Bacterially Expressed Recombinant Coat Protein of Alfalfa mosaic virus

Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1352-1357 ◽  
Author(s):  
B. Khatabi ◽  
B. He ◽  
M. R. Hajimorad
Keyword(s):  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Alfalfa Mosaic Virus ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coat Proteins ◽  
Diagnostic Potential ◽  
Group I ◽  
Wide Range ◽  
Detection And Diagnosis

Alfalfa mosaic virus (AMV), a pathogen of a wide range of plant species, including Glycine max (soybean), is poorly immunogenic. Polyclonal antibodies were generated against bacterially expressed recombinant coat proteins (rCPs) of two biologically distinct AMV strains in rabbits and compared with those raised against native and glutaraldehyde-treated virions of the same strains. Analyses showed that sera against rCPs had comparable antibody titers in indirect enzyme-linked immunosorbent assay with those raised against virions when soybean sap containing homologous viruses served as antigens. Polyclonal antibodies against rCPs were specific, sensitive, and detected all AMV isolates that originated from soybean fields from geographically different regions of the United States. Comparison of CP genes of these isolates showed 96 to 99 and 96 to 100% nucleotide and amino acid sequence identities, respectively, suggesting that they are all closely related. This was further confirmed by phylogenetic analysis where they were all clustered together along with representative AMV strains belonging to group I. Collectively, our data demonstrate that, despite poor immunogenicity of AMV, polyclonal antibodies against rCP are effective probes for detection and diagnosis of the virus.

scholarly journals Evaluation of an Enzyme-Linked Immunosorbent Assay Based on Crude Leishmania Histone Proteins for Serodiagnosis of Human Infantile Visceral Leishmaniasis

2012 ◽  
Vol 19 (9) ◽  
pp. 1487-1491 ◽  
Author(s):  
Sami Lakhal ◽  
Salima Mekki ◽  
Imène Ben-Abda ◽  
Mohamed Mousli ◽  
Fethi Amri ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Immunosorbent Assay ◽  
Diagnostic Performance ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Human Visceral Leishmaniasis ◽  
Diagnostic Potential ◽  
Specificity And Sensitivity ◽  
Nuclear Extracts

ABSTRACTHuman visceral leishmaniasis (VL) is routinely diagnosed by detecting IgG that specifically binds toLeishmaniaantigens. The enzyme-linked immunosorbent assay (ELISA) remains a widely used method. However, the biggest challenge remains the choice of antigen with the highest specificity and sensitivity. This study is aimed at assessing the diagnostic performances of crudeLeishmaniahistone (CLH) protein-based ELISAs in Mediterranean VL patients. The CLH proteins were biochemically purified from promastigote nuclear extracts. Their reactivities were analyzed by Western blotting (WB) using rabbit polyclonal antibodies againstLeishmaniarecombinant histones and sera from VL patients, respectively. Then, the diagnostic potential of CLH proteins was validated by the CLH-based ELISA using 42 infantile VL patients' sera and 70 control subjects. The CLH-based ELISA performance was compared to that of the solubleLeishmaniaantigen (SLA)- and the recombinant K39 (rK39)-based ELISAs. Analysis of the WB profile with the use of polyclonal antibodies confirmed the histone origin of low molecular mass proteins (12 to 16 kDa). All VL samples tested presented antibodies reacting against different antigen fractions; however, recognition patterns were different depending on the reactivity of each serum. CLH-based ELISA showed an excellent ability to discriminate between VL cases and healthy controls (97.6% sensitivity and 100% specificity). It had a diagnostic performance similar to that of rK39-based ELISA (97.6% sensitivity and 97.1% specificity,P= 0.5) and a better serodiagnosis accuracy than the SLA-based ELISA (85.7% sensitivity and 90% specificity,P< 0.05). Therefore, crudeLeishmaniahistone extract could be a valuable antigen for clinical use.

Molecular cloning and partial characterization of a nematode-specific 24 kDa protein fromAscaris suum

Parasitology ◽  
2004 ◽  
Vol 130 (1) ◽  
pp. 131-139 ◽  
Author(s):  
M. K. ISLAM ◽  
T. MIYOSHI ◽  
Y. YOKOMIZO ◽  
N. TSUJI
Keyword(s):  
Amino Acids ◽  
Polyclonal Antibodies ◽  
Protein S ◽  
Immunoblot Analysis ◽  
Toxocara Canis ◽  
Specific Protein ◽  
Reading Frame ◽  
Secretory Products ◽  
Immunohistochemical Studies

The cloning and molecular characterization of a cDNA encodingAscaris suum24 kDa antigen (As24) are described. The cDNA sequence consists of 853 bp with an open reading frame coding for a protein of 147 amino acids with an inferred signal peptide of 19 amino acids. The predicted molecular mass and pI were 16 kDa and 8·35 respectively. The endogenous protein in adultA. suumwas 24 kDa with the expected pI. A search of the public databases revealed over 50% homology with proteins from filarial parasites but not to other known proteins, suggesting that As24 is a nematode-specific protein. Immunohistochemical studies using polyclonal antibodies raised againstEscherichia coli-expressed recombinant As24 demonstrated that the endogenous As24 proteins were intensely localized in unembryonated eggs within the uterus, uterine and gut epithelium, muscle tissues and in the hypodermis of an adult femaleA. suum. Endogenous As24 was expressed throughoutA. suumdevelopment and was detected in the excretory/secretory products by immunoblot analysis. Importantly, a homologous protein(s) was detected inAscarisfrom human andToxocara canisfrom dog, suggesting that As24 is a nematode-specific protein.

Immunocytochemical methodology for the localization of neuronal antigens at the ultrastructural level

1990 ◽  
Vol 48 (3) ◽  
pp. 876-877
Author(s):  
Jorge Pecci Saavedra ◽  
Mark Connaughton ◽  
Juan José López ◽  
Alicia Brusco
Keyword(s):  
Spinal Cord ◽  
Substantia Nigra ◽  
Polyclonal Antibodies ◽  
Ultrastructural Level ◽  
Point Of View ◽  
Nerve Tissue ◽  
Tissue Fixation ◽  
Optimal Conditions ◽  
Interpeduncular Nucleus ◽  
Technical Point

The use of antibodies as labels for the localization of specific molecules in the nervous systan has been extensively applied in recent years. Both monoand polyclonal antibodies or antisera have been employed. The knowledge of the organization of neuronal connectivities, gliovascular relationships, glioneuronal relationships and other features of nerve tissue has greatly increased.A number of areas of the nervous systan have been analyzed in our laboratory, including the nuclei of the raphe system, the reticular formation, interpeduncular nucleus, substantia nigra, caudate nucleus, putamen, pallidum, spinal cord, pineal gland and others.From a technical point of view, a number of variables needed to be taken into account in order to obtain reliable and reproducible results. The design of the optimal conditions of tissue fixation, embedding, sectioning, dilution of antibodies, and adaptation of Sternberger PAP technique were sane of the parameters taken into account to optimize the results. It is critical that each step of the technique be defined for each particular case.

Limitations of dual time point FDG-PET imaging in the evaluation of focal abdominal lesions

2004 ◽  
Vol 43 (05) ◽  
pp. 143-149 ◽  
Author(s):  
N. Hamscho ◽  
C. Menzel ◽  
L. Neuss ◽  
A. F. Kovács ◽  
F. Grünwald ◽  
...  
Keyword(s):  
Pet Imaging ◽  
Malignant Tumors ◽  
Standardized Uptake Value ◽  
Delayed Imaging ◽  
Ct Guided ◽  
Fdg Uptake ◽  
Diagnostic Potential ◽  
Dual Time Point ◽  
Abdominal Lesions ◽  
Time Point

Summary:Aim: For the evaluation of the diagnostic potential of dual time point FDG positron emission tomography (PET) in patients with suspicious focal abdominal up-take, dual time point PET imaging was compared with clinical findings. Patients, methods: In a prospective study, 56 patients exhibiting a solitary suspicious, intense abdominal FDG uptake, underwent dual time point PET imaging for staging or restaging of different malignant tumors, maximal standardized uptake value (SUVmax) measurements included. The first acquisition was started 64.8 ± 19.5, the second 211.3 ± 52.5 min after FDG injection. The final diagnosis based on CT or MRT imaging and a follow-up period of 12.6 ± 2.8 months. Additionally, colonoscopy was done in 6 patients. In another 6 patients histopathology was obtained from CT guided biopsy. Results: Malignant focal abdominal lesions with a SUVmax <2.5 (n = 4) showed an uptake increase of ≥30%. In the remaining malignant cases with an uptake of ≥2.5 (n = 11), up-take increased in 64% and decreased in 36%. Malignant lesions showing FDG uptake decrease (n = 4) had an initial SUVmax value ≥2.5 and remained with a SUVmax ≥2.5 in the second imaging. In benign lesions with an initial SUVmax ≥2.5 (n = 31), the uptake increased in 17 patients (55%) and decreased in 14 patients (45%). All lesions which changed configuration (33%) were confirmed as benign (n = 5). Conclusion: Using dual time point PET abdominal lesions show a very hetergenous uptake pattern regardless of their dignity. Malignancy can only be reliably excluded in lesions which change their configuration and in lesions with an initial SUVmax value <2.5 combined with an SUV decrease in the delayed imaging. Particularly abdominal lesions which show an initial SUVmax ≥2.5 combined with a SUV increase in the delayed imaging are suspicious for malignancy and need further clarification.

Immunological Relationship Between Plasminogen Activator Inhibitors from Different Sources

1987 ◽  
Vol 57 (01) ◽  
pp. 029-034 ◽  
Author(s):  
Göran Urdén ◽  
Joanna Chmielewska ◽  
Tomas Carlsson ◽  
Björn Wiman
Keyword(s):  
Plasminogen Activator ◽  
Polyclonal Antibodies ◽  
Active Form ◽  
Polyclonal Antiserum ◽  
The Other ◽  
Hep G2 ◽  
Inhibitor Activity ◽  
Tissue Plasminogen ◽  
Immunological Relationship ◽  
Different Sources

SummaryPolyclonal antibodies have been raised against the inhibitor moiety in the purified complex between tissue plasminogen activator and its fast inhibitor (PA-inhibitor) in human plasma/ serum. A radioimmunoassay for quantitation of PA-inhibitor antigen was developed. The polyclonal antiserum and a previously described monoclonal antibody against the PA-inhibitor (14) have been used to study the immunological relationship between PA-inhibitors from plasma, serum, platelets, placenta extract and conditioned media from Hep G2 and HT 1080 cells. It was demonstrated that the ratio between PA-inhibitor activity and antigen varied considerably between the different sources. In the plasma samples studied, similar activity and antigen concentrations were found, suggesting that the PA-inhibitor in these samples mainly was in an active form. On the other hand the other sources seemed to contain variable amounts of inactive PA-inhibitor forms. Immunoadsorption experiments revealed that the PA-inhibitor (activity and antigen) from all the sources were specifically bound to the insolubilized antibodies (polyclonal and monoclonal). In no case, however, could active PA-inhibitor be eluted from the immunoadsorption columns. Also the competitive radioimmunoassays suggested that the PA-inhibitors from the different sources studied, were closely immunologically related.

An Immunoradiometric Assay for Factor VIII Related Antigen (VIIIRAg) Using Two Monoclonal Antibodies-Comparison with Polyclonal Rabbit Antibodies for Use in von Willebrand’s Disease Diagnosis

1984 ◽  
Vol 52 (03) ◽  
pp. 250-252 ◽  
Author(s):  
Y Sultan ◽  
Ph Avner ◽  
P Maisonneuve ◽  
D Arnaud ◽  
Ch Jeanneau
Keyword(s):  
Monoclonal Antibodies ◽  
Structural Changes ◽  
Polyclonal Antibodies ◽  
Disease Diagnosis ◽  
Immunoradiometric Assay ◽  
Von Willebrand's Disease ◽  
Von Willebrand’S Disease ◽  
Structural Abnormalities ◽  
Antigenic Material ◽  
Von Willebrand

SummaryTwo monoclonal antibodies raised against FVIII/von Willebrand protein were used in an immunoradiometric assay (IRMA) to measure this antigen in normal plasma and plasma of patients with different forms of von Willebrand’s disease. The first antibody, an IgG1 was used to coat polystyrene tubes, the second one, an IgG2a, iodinated and used in the second step. Both antibodies inhibit ristocetin induced platelet agglutination and react strongly with platelets, megacaryocytes and endothelial cells. The IRMA test using these antibodies showed greater sensitivity than that using rabbit polyclonal anti VIIIRAg antibodies. A good correlation between the two tests was nevertheless found when VIIIRAg was measured in the majority of patient’s plasma. However 5 patients from 3 different families showed more antigenic material in the rabbit antibody IRMA than in the monoclonal antibody IRMA. It is suggested therefore that the monoclonal antibodies identify part of the VIIIR:Ag molecule showing structural abnormalities in these vWd patients, these structural changes remaining undetected by the polyclonal antibodies.

Sign in / Sign up

Export Citation Format

Share Document