Evaluation of reactivity to Echinococcus spp. among rural inhabitants in Poland

2015 ◽  
Vol 60 (3) ◽  
Author(s):  
Ewa Cisak ◽  
Jacek Sroka ◽  
Angelina Wójcik-Fatla ◽  
Violetta Zając ◽  
Jacek Dutkiewicz
Keyword(s):  
Western Blot ◽  
Echinococcus Granulosus ◽  
Cross Reactivity ◽  
Farm Animals ◽  
Control Group ◽  
Laboratory Diagnostics ◽  
Group 14 ◽  
Serum Samples ◽  
The City ◽  
Echinococcus Spp

AbstractA group of 172 rural inhabitants from eastern Poland (68 males and 104 females, mean age 49.0 ± 12.0 years) was examined for the presence of antibodies against Echinococcus granulosus and Echinococcus multilocularis. A population of 38 healthy urban dwellers from the city of Lublin (17 males and 21 females, mean age 36.2 ± 9.6 years) were examined as a control group. Sera of 22 rural inhabitants (12.8%) reacted positively to Echinococcus granulosus hydatid fluid antigen in the screening test. A cross-reactivity was observed with two serum samples that tested positive in ELISA for E. granulosus. Three serum samples were tested positive for E. multilocularis using the Em2plus ELISA assay and also positive for Western blot. None of the members of control group showed the presence of a seropositive reaction to Echinococcus spp. The reactivity to Echinococcus spp. among rural inhabitants decreased with age and this correlation was statistically significant (R = -0.197151, p = 0.009535). The percentage of positive findings was the highest (50.0%) in the youngest age group (14-20). No significant correlations were found between responses to interview questions (possession of domestic and farm animals, contact with wild animals, eating unwashed berries, drinking unboiled water) and the presence of seropositive reactions to Echinococcus spp. The presented results seem to indicate that echinococcosis is still a current problem in Poland that should not be neglected and, moreover, indicates the need for improvement in the routine laboratory diagnostics of Echinococcus spp. by standardizing the ELISA and Western blot tests.

Seroprevalence of toxocariasis in hypereosinophilic individuals in Ahwaz, south-western Iran

2011 ◽  
Vol 86 (2) ◽  
pp. 241-244 ◽  
Author(s):  
S. Maraghi ◽  
A. Rafiei ◽  
R. Hajihossein ◽  
S. M. Sadjjadi
Keyword(s):  
Western Blot ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Infection Frequency ◽  
Serum Samples ◽  
Western Iran ◽  
And Gender ◽  
The City

AbstractEosinophilia in human peripheral blood is caused by different agents, including toxocariasis. The present study aimed to evaluate the prevalence of toxocariasis in hypereosinophilic individuals in the city of Ahwaz, located in south-western Iran, using enzyme-linked immunosorbent assay and Western blot techniques. Serum samples were examined from 100 individuals with peripheral blood eosinophilia and also from another 100 individuals without eosinophilia as the control group. In hypereosinophilic individuals seroprevalence antibodies against Toxocara were found in 19 (19%), of whom 12 (63.15%) were female and 7 (36.85%) were male. Positive sera were subsequently confirmed by Western blot. All of the observed bands ranged from 24 to 100 kDa. Antibodies against Toxocara were found in 1% of the control group, but were not confirmed by Western blot. The results showed significant differences between the frequency of infection within age and gender (P < 0.05); the highest prevalence of infection was observed in adults. Differences between the hypereosinophilic and healthy individuals, in terms of Toxocara infection frequency, also proved significant (P < 0.05).The present study thus confirmed the significant prevalence of toxocariasis as a hygienic problem among hypereosinophilic individuals in this area. It is, therefore, necessary to examine these individuals for toxocariasis.

scholarly journals Maternal Enterovirus Infection during Pregnancy as a Risk Factor in Offspring Diagnosed with Type 1 Diabetes between 15 and 30 Years of Age

2008 ◽  
Vol 2008 ◽  
pp. 1-6 ◽  
Author(s):  
Maria Elfving ◽  
Johan Svensson ◽  
Sami Oikarinen ◽  
Björn Jonsson ◽  
Per Olofsson ◽  
...  
Keyword(s):  
Type 1 Diabetes ◽  
Young Adulthood ◽  
Immunoglobulin M ◽  
Control Group ◽  
Enterovirus Infection ◽  
Group 14 ◽  
Serum Samples ◽  
Specific Immunoglobulin ◽  
Adolescence And Young Adulthood

Maternal enterovirus infections during pregnancy may increase the risk of offspring developing type 1 diabetes during childhood. The aim of this study was to investigate whether gestational enterovirus infections increase the offspring's risk of type 1 diabetes later in life. Serum samples from 30 mothers without diabetes whose offspring developed type 1 diabetes between 15 and 25 years of age were analyzed for enterovirus-specific immunoglobulin M (IgM) antibodies and enterovirus genome (RNA), and compared to a control group. Among the index mothers, 9/30 (30%) were enterovirus IgM-positive, and none was positive for enterovirus RNA. In the control group, 14/90 (16%) were enterovirus IgM-positive, and 4/90 (4%) were positive for enterovirus RNA (n.s.). Boys of enterovirus IgM-positive mothers had approximately 5 times greater risk of developing diabetes (OR 4.63; 95% CI 1.22–17.6), as compared to boys of IgM-negative mothers (P<.025). These results suggest that gestational enterovirus infections may be related to the risk of offspring developing type 1 diabetes in adolescence and young adulthood.

scholarly journals Combined Use of Indirect ELISA and Western Blotting with Recombinant Hepatocellular Carcinoma-Associated Antigen 59 Is a Potential Immunodiagnostic Tool for the Detection of Prepatent Haemonchus contortus Infection in Goat

Animals ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 548 ◽  
Author(s):  
Muhammad Ali-ul-Husnain Naqvi ◽  
Sana Zahra Naqvi ◽  
Muhammad Ali Memon ◽  
Kalibixiati Aimulajiang ◽  
Muhammad Haseeb ◽  
...  
Keyword(s):  
Western Blot ◽  
Western Blotting ◽  
Haemonchus Contortus ◽  
Indirect Elisa ◽  
Fasciola Hepatica ◽  
False Negative ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Combined Use

Haemonchus contortus is recognized as one of the important health problems in small ruminants, leading to reduced production and economic loss for farmers worldwide. Prepatent diagnosis of H. contortus infection is crucial to improve control strategies as this helminth may remove up to one-fifth of total erythrocytes and may cause anemia, edema, diarrhea, and ultimately death in young animals. In this study, one of the excretory and secretory products, rHc-HCA59, was purified and used as antigen to detect specific antibodies in H. contortus infected goats during prepatent stage of infection using indirect enzyme linked immunosorbent assay (ELISA) as screening test. All goats (n = 38) were housed indoor, experimentally infected with 8000 infective larvae (L3) of H. contortus, and serum samples were collected prior to infection and at 14th day of infection. Immunoblotting was performed to confirm the results of indirect ELISA, evaluate the cross reactivity against rHc-HCA59 in sera of most common co-infecting parasites and rectify the false negative samples. Furthermore, three different batches of rHc-HCA59 were produced to evaluate the repeatability of ELISA. No eggs were detected in feces of all goats collected at 7th and 14th day of infection but, H. contortus eggs were detected at 21 days post infection in the feces. Indirect ELISA performed in this study showed 87% sensitivity and 100% specificity. The western blot analysis confirmed immunoreactivity in serum samples which scored positive in indirect ELISA and recognized the samples as negative which had OD450 lower than negative cut-off value in indirect ELISA. Furthermore, all false negative sera (n = 5) that had OD450 value between positive and negative cut-off value in rHc-HCA59 based ELISA were clearly positive in western blot. Moreover, no cross-reactivity was detected in ELISA and western blotting against rHc-HCA59 in positive sera of Toxoplasma gondii, Fasciola hepatica, and Trichinella spiralis. The results of this study concluded that combined use of indirect ELISA and western blotting with rHc-HCA59 is a potential immunodiagnostic tool for the detection of H. contortus infection during prepatent period in goats.

scholarly journals Increase of Angiogenesis and  Osteogenesis  by Local Application of Simvastatin Depends on VEGF

2020 ◽  
Author(s):  
Jie Tan ◽  
Hong Wang ◽  
Guohong Du ◽  
Huijie Leng ◽  
Chunli Song
Keyword(s):  
Bone Formation ◽  
Western Blot ◽  
Dual Energy ◽  
Male Rats ◽  
Control Group ◽  
Sprague Dawley ◽  
Micro Computed Tomography ◽  
Vegf Expression ◽  
Serum Samples ◽  
X Ray

Abstract Aims Simvastatin stimulates both BMP-2 and VEGF expression, but it is unknown which is more important for bone formation. This study was undertaken to determine whether these effects could be blocked by the anti-VEGF antibody bevacizumab. Methods 60 Sprague–Dawley male rats were randomly divided into five groups (n = 12 per group): normal control; simvastatin 0 mg or 0.5 mg; and bevacizumab with simvastatin 0 mg or 0.5 mg. Simvastatin groups were administered intraosseous injections of simvastatin delivered by thermosensitive poloxamer. Bevacizumab groups were given bevacizumab intraperitoneally or the same volume of saline. Serum samples were collected before the treatment and every two weeks thereafter. Four weeks after the treatment, four rats randomly selected from each group were subjected to Microfil® perfusion. The remaining eight rats was evaluated with dual energy X-ray absorptiometry (DXA) and micro-computed tomography (µCT). Four specimens (left tibias) were randomly selected from each group for undecalcified histology, the other four specimens selected for Western blot to analyse the changes of expression of BMP-2. Results local injection of simvastatin significantly increased bone formation. Microfil® perfusion showed that there were more vessels both in the bone marrow and around the bone after a single-dose simvastatin injection. Western blot analysis also confirmed that the expression levels of BMP2 were significantly higher in the simvastatin-treated groups compared with the control group. Compared with the simvastatin group, bevacizumab blunted the simvastatin-induced increase in bone mass and angiogenesis. Conclusion The anabolic effect of simvastatin on bone formation is through VEGF-related mechanisms.

scholarly journals Western blot assay of anti-Echinococcus granulosus antibody positive serum samples by indirect haemagglutination method

2019 ◽  
Vol 76 (2) ◽  
pp. 195-202 ◽  
Author(s):  
Ayşe Semra Güreser ◽  
Gamze Gizem Duman ◽  
Fakhriddin Sarzhanov ◽  
Djursun Karasartova ◽  
Funda Dogruman Al ◽  
...  
Keyword(s):  
Western Blot ◽  
Echinococcus Granulosus ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Indirect Haemagglutination ◽  
Positive Serum

scholarly journals Development of antibodies to pan-coronavirus spike peptides in convalescent COVID-19 patients

2020 ◽  
Author(s):  
Andrii Rabets ◽  
Galyna Bila ◽  
Roman Grytsko ◽  
Markian Samborsky ◽  
Yuriy Rebets ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Membrane Surface ◽  
Protein S ◽  
Cellular Membrane ◽  
Cross Reactivity ◽  
Protein Domain ◽  
Spike Protein ◽  
Heptad Repeat ◽  
Control Group ◽  
Serum Samples

Coronaviruses are sharing several protein regions notable the spike protein (S) on their enveloped membrane surface, with the S1 subunit recognizing and binding to the cellular receptor, while the S2 subunit mediates viral and cellular membrane fusion. This similarity opens the question whether infection with one coronavirus will confer resistance to other coronaviruses? Investigating patient serum samples after SARS-CoV-2 infection in cross-reactivity studies of immunogenic peptides from Middle East respiratory syndrome coronavirus (MERS-CoV), we were able to detect the production of antibodies also recognizing MERS virus antigens. The cross-reactive peptide comes from the heptad repeat 2 (HR2) domain of the MERS virus spike protein. Indeed, the peptide of the HR2 domain of the MERS spike protein, previously proven to induce antibodies against MERS-CoV is sharing 74% homology with the corresponding sequence of SARS-CoV-19 virus. Sera samples of 47 convalescent SARS-CoV-2 patients, validated by RT-PCR-negative testes 30 days post-infection, and samples of 40 sera of control patients (not infected with SARS-CoV-2 previously) were used to establish eventual cross-bind reactivity with the MERS peptide antigen. Significantly stronger binding (p<0.0001) was observed for IgG antibodies in convalescent SARS-CoV-2 patients compared to the control group. If used as an antigen, the peptide of the HR2 domain of the MERS spike protein allows discrimination between post-Covid populations from non-infected ones by the presence of antibodies in blood samples. This suggests that polyclonal antibodies established during SARS-CoV-2 infection has the ability to recognize and probably decrease infectiveness of MERS-CoV infections as well as other coronaviruses. The high homology of the spike protein domain suggests in addition that the opposite effect can also be true: coronaviral infections producing cross-reactive antibodies affective against SARS-CoV-19. The collected data prove in addition that despite the core HR2 region being hidden in the native viral conformation, its exposure during cell entry makes it highly immunogenic. Since inhibitory peptides to this region were previously described, this opens new possibilities in fighting coronaviral infections.

scholarly journals Assessment of serological tests for antibodies to different antigens of the SARS-CoV-2 virus: comparison of six immunoassays

2021 ◽  
Vol 23 (6) ◽  
pp. 1395-1404
Author(s):  
V. V. Belyakova ◽  
O. A. Maiorova ◽  
N, V. Ivanova ◽  
I. E. Stepanova ◽  
M. A. Smerdova ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Rapid Test ◽  
Diagnostic Methods ◽  
Federal State ◽  
Control Group ◽  
Laboratory Diagnostics ◽  
Igg Antibodies ◽  
Diagnostic Systems ◽  
Serum Samples ◽  
Serological Tests

The new coronavirus SARS-CoV-2 has become a global challenge to medicine and, in particular, laboratory diagnostics. The study of the antibodies’ level to SARS-CoV-2 can be used as a confirmation test in the diagnosis of a disease, but it becomes of paramount importance in assessing population immunity resulting from a disease or vaccination, as well as in selection of convalescent plasma donors. The kits developed in our country and abroad for detecting antibodies to the SARS-CoV-2 virus differ both in the methods of testing and in the used coronavirus antigens to which the antibodies are directed. The aim of this study was to compare the diagnostic sensitivity and specificity of five kits for the detection of IgG antibodies to the SARS-CoV-2 virus, based on different diagnostic methods. Serum samples from 137 COVID-19 convalescents and 166 donors of blood and its components were examined. The control group consisted of 50 blood sera collected at the beginning of 2019 and 19 sera collected in 2018 (before the advent of the SARS-CoV-2 virus) and stored at -70 °C. Testing was carried out in analytical systems: rapid test “COVID-19 IgM/IgG Rapid Test (Colloidal Gold)” (China), on an automatic immunochemical analyzer Abbott Architect™ i2000 and kit “SARS-CoV2-IgG” (Abbot, Chicago , IL USA), by the chemiluminescence method using an automatic analyzer of the CL series and kits of the “Mindray” company (China) “SARS-CoV-2 IgM” and “SARS-CoV-2 IgG” and by the enzyme immunoassay method on the kits of the companies “Diagnostic Systems” Ltd (Russia, Nizhny Novgorod) “DS-IFA-ANTI-SARS-CoV-2-G”, “Xema” Ltd (Federal State Budgetary Institution “National Medical Research Center of Hematology” of the Ministry of Health of Russia) “SARS-CoV-2-IgG-IFA” and “Vector-Best” CJSC (Russia, Novosibirsk)” SARS-COV-2-IgM-IFA-BEST” and “SARS-COV-2-IgG-IFABEST”. When comparing the results of testing 137 plasma samples on the Vector-Best and Mindray kits for IgG antibodies, 127 samples were positive, 7 samples were negative on both kits, the discrepancy was 2.2%. In the study of IgM antibodies, 32.1% were positive, and 52.6% were negative in both kits. The discrepancy rate was 15.3%. Out of 166 samples, 1 serum (0.6%) was negative in 5 kits. On the Mindray kit, IgG antibodies to the antigens of the SARS-CoV-2 virus were detected in 165 samples (99.4%), on Vector-Best – in 164 sera (98.8%), on Diagnostic systems – in 151 (90.96%), on Xema – in 154 (92.8%), and on Abbott – in 155 samples (93.4%). At the same time, 135 (81.33%) samples were positive in all kits, while 30 samples had discordant results (18.07%), and in 9 sera, specific IgG was not detected in 2 or more kits. ROC analysis revealed a high diagnostic value of all tested kits (AUC from 0.908 to 0.998), which indicates a high quality of the separation model of positive and negative samples (p < 0.001). With the cut-off set by the manufacturers, the sensitivity and specificity ranged from 82.8% and 93.3% for the Diagnostic Systems kit to 99.4% and 95.8% for the VectorBest kit. The calculated correlation coefficients were higher between kits with a similar composition of the antigen used in the kits; therefore, it is better to monitor the dynamics of antibodies by diagnostic kits from the same manufacturer.

scholarly journals Western Immunoblotting for Bartonella Endocarditis

2003 ◽  
Vol 10 (1) ◽  
pp. 95-102 ◽  
Author(s):  
Pierre Houpikian ◽  
Didier Raoult
Keyword(s):  
Western Blot ◽  
Cross Reactivity ◽  
Molecular Techniques ◽  
Cat Scratch Disease ◽  
Serum Samples ◽  
Western Immunoblotting ◽  
Serological Tests ◽  
Heterologous Antigen ◽  
Bartonella Quintana ◽  
Causative Species

ABSTRACT To differentiate infectious endocarditis (IE) from other Bartonella infections and to identify infecting Bartonella bacteria at the species level on a serological basis, we used Western immunoblotting to test sera from 51 patients with Bartonella IE (of which 27 had previously benefited from species identification by molecular techniques), 11 patients with chronic Bartonella quintana bacteremia, and 10 patients with cat scratch disease. Patients with IE were Western blot positive in 49 of 51 cases, and significant cross-reactivity with three heterologous Bartonella antigens was found in 45 of 49 cases. Sera from bacteremic patients did not react with more than one heterologous antigen, and sera from patients with cat scratch disease gave negative results. Sera reacted only with B. henselae in four cases of IE, including one with a positive PCR result for valve tissue. Western blot and cross-adsorption performed on serum samples from patients with IE (the identity of the causative species having been determined by PCR) were demonstrated to identify efficiently the causative species in all cases. When applied to patients diagnosed on the basis of serological tests only, this technique allowed identification of the causative species in 20 of 22 cases. The results were in accordance with epidemiological features. Six reactive bands of B. quintana (of molecular sizes from 10 to 83 kDa) demonstrated significant association with sera from patients with B. quintana endocarditis. Overall, Western blotting and cross-adsorption made it possible to identify the causative species in 49 of 51 (96%) IE cases.

scholarly journals Is the prevalence of celiac disease increased among epileptic patients?

2003 ◽  
Vol 61 (2B) ◽  
pp. 330-334 ◽  
Author(s):  
Riccardo Pratesi ◽  
Lenora Gandolfi ◽  
Rita C. Martins ◽  
Pedro L. Tauil ◽  
Yanna Karla Nobrega ◽  
...  
Keyword(s):  
Small Intestine ◽  
Celiac Disease ◽  
Clinical Analysis ◽  
Control Group ◽  
Serum Samples ◽  
Blood Testing ◽  
General Hospitals ◽  
Epileptic Patients ◽  
The City ◽  
Group 1

OBJECTIVE: To assess the prevalence of celiac disease (CD) among a group of epileptic patients attending the Epilepsy Clinics of two general hospitals in the city of Brasilia (DF), Brazil. METHOD: Serum samples were collected from 255 epileptic patients (119 children, 136 adults) originating from Epilepsy Clinics, and from a control group composed by 4405 individuals (2034 children, 2371 adults) attending the Laboratory of Clinical Analysis, for routine blood testing. The diagnosis of CD was determined by the antiendomysium antibody (IgA-EMA) test and by small intestine biopsy. RESULTS: two of the 255 epileptic patients (1:127) and fifteen subjects from the control group (1:293) tested positive for the IgA-EMA assay. CONCLUSION: the prevalence of CD was 2.3 times higher in epileptic patients than in controls (7.84 per 1000 versus 3.41 per 1000). Although still not statistically significant, this result is highly suggestive of an increased prevalence of CD among epileptic patients.

scholarly journals False-Positive Results in a Recombinant Severe Acute Respiratory Syndrome-Associated Coronavirus (SARS-CoV) Nucleocapsid-Based Western Blot Assay Were Rectified by the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV

2006 ◽  
Vol 13 (3) ◽  
pp. 409-414 ◽  
Author(s):  
Mimoun Maache ◽  
Florence Komurian-Pradel ◽  
Alain Rajoharison ◽  
Magali Perret ◽  
Jean-Luc Berland ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Western Blot ◽  
False Positive ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Western Blot Assay ◽  
N Protein ◽  
False Positive Diagnosis ◽  
Healthy Donors

ABSTRACT To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV.

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