scholarly journals Identification of toxocara canis antigens by Western blot in experimentally infected rabbits

2002 ◽  
Vol 44 (4) ◽  
pp. 213-216 ◽  
Author(s):  
Olga Lucía MORALES ◽  
Myriam Consuelo LÓPEZ ◽  
Rubén Santiago NICHOLLS ◽  
Carlos AGUDELO
Keyword(s):  
Western Blot ◽  
Toxocara Canis ◽  
Diagnostic Techniques ◽  
Elisa Test ◽  
Elisa Assay ◽  
Secondary Antibody ◽  
Igg Antibodies ◽  
Specific Antibodies ◽  
Second Stage ◽  
Specific Antigens

Toxocariasis is a frequent helminthiasis that can cause visceral and ocular damage in humans specially in children. The identification of specific antigens of Toxocara canis is important in order to develop better diagnostic techniques. Ten rabbits were infected orally with a dose of 5000 Toxocara canis embryonated eggs. Rabbits were bled periodically and an ELISA assay was performed to determine levels of specific Toxocara IgG antibodies. ELISA detected antibodies at day 15 after infection. Western blot (WB) assay was performed using excretory/secretory antigens (E/S) of T. canis second stage larvae. Different antigen concentrations were evaluated: 150, 200, 250 and 300 µg/mL. The concentration of 250 µg/mL was retained for analysis. Rabbit sera were diluted 1:100. Secondary antibody was used at a dilution of 1:1000. Results of WB indicated that in the first month after infection specific antibodies against the 200 KDa, 116 KDa, 92 KDa and 35 KDa antigens were detected; antibodies against the 92 KDa, 80 KDa, 66 KDa, 45 KDa, 31 KDa and 28 KDa antigens appeared later. All positive sera in the ELISA test were also positive in WB. Two antigen bands, 92 KDa and 35 KDa, were identified since the beginning and throughout the course of infection. These antigens merit further evaluation as candidates for use in diagnosis.

scholarly journals Effect of multiple freeze-thaw cycles on detection of anti-SARS-CoV-2 IgG antibodies

2021 ◽  
Author(s):  
Farah M. Shurrab ◽  
Duaa W. Al-Sadeq ◽  
Fathima Amanullah ◽  
Salma N. Younes ◽  
Hadeel Al-Jighefee ◽  
...  
Keyword(s):  
Random Effect ◽  
Elisa Test ◽  
Antibody Detection ◽  
P Value ◽  
Elisa Assay ◽  
Igg Antibodies ◽  
Rna Detection ◽  
Freeze Thaw ◽  
Significant Difference ◽  
Number Of Cycles

AbstractSeveral studies have investigated the effect of repeated freeze-thaw (F/T) cycles on RNA detection for SARS-CoV-2. However, no data is available regarding the effect of repeated F/T cycles on SARS-CoV-2 antibody detection in serum. We investigated the effect of multiple F/T cycles on anti-SARS-CoV-2 IgG detection using an ELISA test targeting the nucleocapsid antibodies. Ten positive and one negative SARS-CoV-2 IgG sera from 11 participants, in replicates of five were subjected to a total of 16 F/T cycles and stored at 4°C until tested by ELISA. Statistical analysis was done to test for F/T cycle effect. Non-of the 10 positive sera turned into negative after 16 F/T cycles. There was no significant difference in the OD average reading between the first and last F/T cycles, except for one serum with a minimal decline in the OD. The random-effect linear regression of log (OD) on the number of cycles showed no significant trend with a slope consistent with zero (B=-0.0001; 95% CI −0.0008; 0.0006; p-value=0.781). These results suggest that multiple F/T cycles had no effect on the ability of the ELISA assay to detect the SARS-CoV-2 IgG antibodies.

Immunocharacterization ofTaenia soliumoncosphere and metacestode antigens

1996 ◽  
Vol 70 (4) ◽  
pp. 271-280 ◽  
Author(s):  
C. Garcia-Allan ◽  
N. Martínez ◽  
A. Flisser ◽  
A. Aluja ◽  
J.C. Allan ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Western Blot ◽  
Experimental Infection ◽  
Blot Analysis ◽  
Post Infection ◽  
Western Blot Analysis ◽  
Taenia Solium ◽  
Specific Antibodies ◽  
Specific Antigens ◽  
Related Increase

AbstractA partial immunocharacterization of oncosphere and metacestode antigens ofTaenia soliumwas carried out and compared to antigens from other taeniid species. The results indicated thatT. soliummetacestode antigen contained epitopes cross reactive with rabbit anti-sera to adult and oncospheral stages of the parasite. Oncospheres, however, consisted largely of stage specific antigens. Western blot analysis indicated thatT. soliumandT. pisiformisshared several oncospheral antigens; however, this was not the case withT. soliumandT. hydatigena. Western blot analysis showed a time-related increase in the number of molecules recognized by antibodies toT. soliumoncosphere and metacestode antigens in pigs experimentally infected withT. soliumeggs. Oncosphere specific antibodies were detected in pig sera one month after experimental infection whereas antibodies to cystic stage antigens were not present until the 3rd to 5th month post infection. Sera from neurocysticercotic patients as well as naturally infected cysticercotic pigs recognized high molecular weight antigens in the oncospheres.

scholarly journals Effect of multiple freeze–thaw cycles on the detection of anti-SARS-CoV-2 IgG antibodies

2021 ◽  
Vol 70 (8) ◽  
Author(s):  
Farah M. Shurrab ◽  
Duaa W. Al-Sadeq ◽  
Fathima Amanullah ◽  
Salma N. Younes ◽  
Hadeel Al-Jighefee ◽  
...  
Keyword(s):  
Statistical Analysis ◽  
Random Effect ◽  
Elisa Test ◽  
Antibody Detection ◽  
Elisa Assay ◽  
Igg Antibodies ◽  
Rna Detection ◽  
Freeze Thaw ◽  
Significant Difference ◽  
Number Of Cycles

Several studies have investigated the effect of repeated freeze–thaw (F/T) cycles on RNA detection for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). However, no data are available regarding the effect of repeated F/T cycles on SARS-CoV-2 antibody detection in serum. We investigated the effect of multiple F/T cycles on anti-SARS-CoV-2 IgG detection using an ELISA test targeting the nucleocapsid antibodies. Ten positive and 1 negative SARS-CoV-2 IgG sera from 11 participants, in replicates of 5, were subjected to a total of 16 F/T cycles and stored at 4 °C until tested by ELISA. Statistical analysis was performed to test for F/T cycle effect. None of the 10 positive sera became negative after 16 F/T cycles. There was no significant difference in the OD average reading between the first and last F/T cycles, except for one serum with a minimal decline in the OD. The random effect linear regression of log (OD) on the number of cycles showed no significant trend, with a slope consistent with zero (B=−0.0001; 95 % CI −0.0008; 0.0006; P-value=0.781). These results suggest that multiple F/T cycles had no effect on the ability of the ELISA assay to detect SARS-CoV-2 IgG antibodies.

scholarly journals Seroprevalence of Toxocara canis in the city of Catania, Italy.

2019 ◽  
Vol 11 (1) ◽  
pp. e2019031 ◽  
Author(s):  
Alessandra Nicoletti ◽  
Calogero Edoardo Cicero ◽  
Antonia Mantella ◽  
Loretta Giuliano ◽  
Cristina Rascunà ◽  
...  
Keyword(s):  
Risk Factors ◽  
Multivariate Analysis ◽  
Urban Areas ◽  
Southern Italy ◽  
Positive Association ◽  
Toxocara Canis ◽  
Elisa Test ◽  
Igg Antibodies ◽  
Face To Face ◽  
The City

Toxocariasis is one of the most common helminthiases worldwide. However, there is a lack of data regarding Southern Italy. We have evaluated the seroprevalence and associated environmental factors of toxocariasis in a sample of adults living in the city of Catania. Presence of anti-Toxocara canis IgG antibodies was searched using an ELISA test using excretory/secretory antigens. Environmental risk factors have beene valuated with a face-to-face questionnaire. Two hundred eighty-seven subjects (mean age of 48.1±15.6 years) were enrolled, and presence of anti T. canis antibodies was found in 23 participants, of whom 18 (78.3%) were women with a mean age of 51.1±14.0 years, giving a seroprevalence of 8.0% (95%CI 5.4-11.7). At multivariate analysis a positive association for subjects with more than 3 siblings (adjOR 3.17; 95%CI 1.09-9.25) was recorded. Our study confirms that exposition to T. canis is common also in urban areas of western countries.

Prevalence of Toxoplasma gondii and Toxocara canis among Myositis Patients in Southwest of Iran

2019 ◽  
Vol 20 ◽  
Author(s):  
Jasem Saki ◽  
Karim Mowla ◽  
Reza Arjmand ◽  
Forough Kazemi ◽  
Somayeh Fallahizadeh
Keyword(s):  
Toxoplasma Gondii ◽  
Nested Pcr ◽  
Healthy Subjects ◽  
Toxocara Canis ◽  
Control Group ◽  
Igg Antibodies ◽  
Healthy Individuals ◽  
High Prevalence ◽  
Acute Toxoplasmosis ◽  
Southwest Of Iran

Introduction: Parasitic myositis is caused by some parasites such as T. gondii and T. canis. So, the aim of the study was to evaluate the prevalence T. gondii and T. canis in patients with myositis and healthy individuals. Methods: A total of 108 samples were randomly selected as the control (54 healthy individuals) and test (54 myositis patients) groups. IgG and IgM antibodies against T. gondii and IgG antibodies against T. canis were measured by the ELISA. The detection of chronic and acute toxoplasmosis was performed by the ELISA IgG avidity. The presence of T. gondii in blood was evaluated by the nested-PCR. Results: Of 108, 33 (30.6%) cases were detected positive for IgG against T. gondii that 19 (35.2%) and 14 (25.9%) were observed in myositis patients and healthy individuals, respectively (P=0.296). Of 19 positive cases, 12 (63.2%) and 7 (36.8%) cases were detected as chronic and acute toxoplasmosis, respectively, while, all positive cases in the control group had chronic toxoplasmosis (P=0.013). One (1.9%) sample was detected positive for anti- Toxoplasma gondii IgM and two (3.7%) samples were found positive for IgG against T. canis by the ELISA that these positive cases were observed only in myositis patients (P=1.000 P=0.495, respectively). B1 T. gondii gene was amplified in 12 (63.2%) and 1 (7.1%) in myositis patients and healthy subjects (P=0.001). Conclusions: Our findings showed that there was a relatively high prevalence of acute toxoplasmosis in myositis patients in comparison with the control subjects in southwest of Iran.

scholarly journals Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

1992 ◽  
Vol 34 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Eide Dias Camargo ◽  
Paulo Mutuko Nakamura ◽  
Adelaide José Vaz ◽  
Marcos Vinícius da Silva ◽  
Pedro Paulo Chieffi ◽  
...  
Keyword(s):  
Low Cost ◽  
Clinical Signs ◽  
Toxocara Canis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Normal Individuals ◽  
Before And After ◽  
Dot Elisa ◽  
Stability Time

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

scholarly journals Mobility Inhibition and Nematocidal Activity of Asarone and Related Phenylpropanoids on Second-Stage Larvae of Toxocara canis.

1995 ◽  
Vol 18 (4) ◽  
pp. 605-609 ◽  
Author(s):  
Naoki SUGIMOTO ◽  
Yoshihisa GOTO ◽  
Nobuaki AKAO ◽  
Fumiyuki KIUCHI ◽  
Kaoru KONDO ◽  
...  
Keyword(s):  
Toxocara Canis ◽  
Second Stage ◽  
Nematocidal Activity

Toxocariasis/cysticercosis seroprevalence in a long-term rural settlement, São Paulo, Brazil

Parasitology ◽  
2009 ◽  
Vol 136 (6) ◽  
pp. 681-689 ◽  
Author(s):  
L. E. PRESTES-CARNEIRO ◽  
D. H. P. SOUZA ◽  
G. C. MORENO ◽  
C. TROIANI ◽  
V. SANTARÉM ◽  
...  
Keyword(s):  
Western Blot ◽  
Taenia Solium ◽  
Sao Paulo ◽  
Rural Settlement ◽  
São Paulo ◽  
Educational Levels ◽  
Second Stage ◽  
Paulo State ◽  
Ocular Manifestations

SUMMARYSeroprevalence of Toxocara and Taenia solium and risk factors for infection with these parasites were explored in a long-term rural settlement in São Paulo state, Brazil. An ELISA for the detection of anti-Toxocara IgG and IgE and anti-T. solium cysticerci was standardized using Toxocara excretory-secretory antigens (TES) obtained from the cultured second-stage larvae of T. canis and by vesicular fluid antigen from Taenia crassiceps cysticerci (VF). For cysticercosis, the reactive ELISA samples were assayed by Western blot using 18 kDa and 14 kDa proteins purified from VF. Out of 182 subjects, 25 (13·7%) presented anti-Toxocara IgG and a positive correlation between total IgE and the reactive index of specific anti-TES IgE (P=0·0265) was found amongst the subjects found seropositive for anti-Toxocara IgG. In these individuals 38·0% showed ocular manifestations. The frequency of anti-T. solium cysticerci confirmed by Western blot was 0·6%. Seropositivity for Toxocara was correlated with low educational levels and the owning of dogs. Embryonated eggs of Toxocara spp. were found in 43·3% of the analysed areas.

scholarly journals Broadly Neutralizing Hemagglutinin Stalk-Specific Antibodies Induce Potent Phagocytosis of Immune Complexes by Neutrophils in an Fc-Dependent Manner

mBio ◽  
2016 ◽  
Vol 7 (5) ◽  
Author(s):  
Caitlin E. Mullarkey ◽  
Mark J. Bailey ◽  
Diana A. Golubeva ◽  
Gene S. Tan ◽  
Raffael Nachbagauer ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Fc Receptor ◽  
Igg Antibodies ◽  
Specific Antibodies ◽  
Effector Functions ◽  
Iga Antibodies ◽  
Specific Igg ◽  
Optimal Protection ◽  
Specific Igg Antibodies

ABSTRACTBroadly neutralizing antibodies that recognize the conserved hemagglutinin (HA) stalk have emerged as exciting new biotherapeutic tools to combat seasonal and pandemic influenza viruses. Our general understanding of the mechanisms by which stalk-specific antibodies achieve protection is rapidly evolving. It has recently been demonstrated that broadly neutralizing HA stalk-specific IgG antibodies require Fc-Fcγ receptor (FcγR) interactions for optimal protectionin vivo. Here we examine the neutrophil effector functions induced by stalk-specific antibodies. As the most abundant subset of blood leukocytes, neutrophils represent a critical innate effector cell population and serve an instrumental role in orchestrating downstream adaptive responses to influenza virus infection. Yet, the interplay of HA stalk-specific IgG, Fc-FcγR engagement, and neutrophils has remained largely uncharacterized. Using anin vitroassay to detect the production of reactive oxygen species (ROS), we show that human and mouse monoclonal HA stalk-specific IgG antibodies are able to induce the production of ROS by neutrophils, while HA head-specific antibodies do not. Furthermore, our results indicate that the production of ROS is dependent on Fc receptor (FcR) engagement and phagocytosis. We went on to assess the ability of monoclonal HA stalk-specific IgA antibodies to induce ROS. Consistent with our findings for monoclonal IgGs, only HA stalk-specific IgA antibodies elicited ROS production by neutrophils. This induction is dependent on the engagement of FcαR1. Taken together, our findings describe a novel FcR-dependent effector function induced by HA stalk-specific IgG and IgA antibodies, and importantly, our studies shed light on the mechanisms by which HA stalk-specific antibodies achieve protection.IMPORTANCEThe present study provides evidence that broadly neutralizing HA stalk-specific antibodies induce downstream Fc-mediated neutrophil effector functions. In addition to their ability to neutralize, this class of antibodies has been shown to rely on Fc-Fc receptor interactions for optimal protectionin vivo. Curiously, neutralizing antibodies that bind the HA head domain do not require such interactions. Our findings build on these previous observations and provide a more complete picture of the relationship between stalk-specific antibodies and cells of the innate immune compartment. Furthermore, our data suggest that the ability of HA stalk-specific antibodies to mediate Fc-Fc receptor engagement is epitope dependent. Overall, this work will inform the rational design of improved influenza virus vaccines and therapeutics.

scholarly journals Comparison between the efficiency of serological tests for Identification of Brucellosis

2010 ◽  
Vol 7 (2) ◽  
pp. 901-909
Author(s):  
Baghdad Science Journal
Keyword(s):  
Rose Bengal ◽  
Serological Test ◽  
Elisa Test ◽  
Health Centers ◽  
Confirmatory Test ◽  
Igg Antibodies ◽  
Serological Tests ◽  
Primary Screening ◽  
Low Concentrations ◽  
Immune Assay

Five serological methods for detection of Brucella were compaired in this study, Four of the methods are commonely used in the detections:- 1-Rose-Bengal: as primary screening test which depends on detecting antibodies in the blood serum. 2-IFAT: which detects IgG and IgM antibodies in the serum. 3-ELISA test: which detects IgG antibodies in the serum. 4-2ME test: which detects IgG antibodies The fifth methods. It was developed by a reasercher in one of the health centers in Baghdad. It was given the name of spot Immune Assay (SIA). Results declares that among (100) samples of patients blood, 76, 49, 49, 37, and 28. samples were positive to Rose Bengal, ELISA, SIA, 2ME and IFAT tests, respectively. When efficiency, sensitivity and specificity of the serological methods were compaired, the Following results were obtained: a) ELISA and SIA were superiors among the other confirming methods (2ME and IFAT) in detecting the highest cases (49 cases); 46 of them were from the (76) cases positive to Rose Bengal The confirmatory test 2ME was not efficient in detecting low concentrations of IgG antibodies when less than half (37) of the total positive cases (76) were detected by this test. b) IFAT test was the least efficient confirmatory test among all other test. c) As a new confirmatory test, SIA proved to be an efficient and serological test for Brucella detection in comparison with other tests. It is an easy to use test, rapid and could be performed without need to the expensive equipment .

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