scholarly journals Seroprevalence Study of Human Brucellosis by Conventional Tests and Indigenous Indirect Enzyme-Linked Immunosorbent Assay

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Annapurna S. Agasthya ◽  
Srikrishna Isloor ◽  
Prabhudas Krishnamsetty
Keyword(s):  
Joint Pain ◽  
Indirect Elisa ◽  
Clinical Signs ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Serum Samples ◽  
Heart Attacks ◽  
Highly Sensitive ◽  
Monitoring And Surveillance

Brucellosis is one of the most important reemerging zoonoses in many countries. Brucellosis is caused by Gram-negative coccobacillus belonging to genusBrucella. Human brucellosis often makes the diagnosis difficult. The symptoms and clinical signs most commonly reported are fever, fatigue, malaise, chills, sweats headaches, myalgia, arthralgia, and weight loss. Some cases have been presented with only joint pain, lower backache, and involuntary limb movement, burning feet, or ischemic heart attacks. The focus of this work was to develop a highly sensitive and specific indirect ELISA by using smooth lipopolysaccharide antigen ofBrucella abortus 99to detect anti-Brucellaantibodies at Project Directorate on Animal Disease Monitoring and Surveillance. Serum samples collected from 652 individuals in whom fever was not the major symptom but the complaint was of joint pain, headache, lower backache, and so forth, were screened by Rose Bengal plate agglutination test (RBPT) and standard tube agglutination test (STAT). Subsequent testing of sera by indigenous indirect ELISA detected 20 samples positive (3.6% seroprevalence), and indirect ELISA was found to be more sensitive than RBPT and STAT. The seroprevalence in South Karnataka was 2.14%, and in North Karnataka it was 0.92%.

scholarly journals Development of a Highly Sensitive and Specific Enzyme-Linked Immunosorbent Assay Based on Recombinant Matrix Protein for Detection of Avian Pneumovirus Antibodies

2000 ◽  
Vol 38 (11) ◽  
pp. 4010-4014 ◽  
Author(s):  
Baldev R. Gulati ◽  
Kjerstin T. Cameron ◽  
Bruce S. Seal ◽  
Sagar M. Goyal ◽  
David A. Halvorson ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Matrix Protein ◽  
Clinical Signs ◽  
M Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Test ◽  
Avian Pneumovirus ◽  
Highly Sensitive ◽  
The Matrix

The matrix (M) protein of avian pneumovirus (APV) was evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (ELISA) for diagnosis of APV infection, a newly emergent disease of turkeys in United States. Sera from APV-infected turkeys consistently contained antibodies to a 30-kDa protein (M protein). An ELISA based on recombinant M protein generated in Escherichia coli was compared with the routine APV ELISA that utilizes inactivated virus as antigen. Of 34 experimentally infected turkeys, 33 (97.1%) were positive by M protein ELISA whereas only 18 (52.9%) were positive by routine APV ELISA 28 days after infection. None of the serum samples from 41 uninfected experimental turkeys were positive by M protein ELISA. Of 184 field sera from turkey flocks suspected of having APV infection, 133 (72.3%) were positive by M protein ELISA whereas only 99 (53.8%) were positive by routine APV ELISA. Twelve serum samples, which were negative by M protein ELISA but positive by routine APV ELISA, were not reactive with either recombinant M protein or denatured purified APV proteins by Western analysis. This indicates that the samples had given false-positive results by routine APV ELISA. The M protein ELISA was over six times more sensitive than virus isolation (11.5%) in detecting infections from samples obtained from birds showing clinical signs of APV infection. Taken together, these results show that ELISA based on recombinant M protein is a highly sensitive and specific test for detecting antibodies to APV.

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.

Sero - Prevalence and Risk Factors of Cattle and Human Brucellosis at Seka Chokorsa and Shebe Sonbo Jimma Zone Districts, South West Ethiopia

2019 ◽  
Author(s):  
Ftsum Assefa Tokon ◽  
Benti Deresa Gelalcha ◽  
Teferi Benti Moti ◽  
Redeat Belaneh Alemu ◽  
Hailu Degefu Awash
Keyword(s):  
Public Health ◽  
Risk Factors ◽  
Brucella Abortus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Bacterial Disease ◽  
Serum Samples ◽  
Public Health Importance ◽  
Herd Level ◽  
Jimma Zone

Abstract Background: Brucellosis is contagious bacterial disease of major socio-economic and public health importance globally and it is also one of the priority zoonotic diseases in Ethiopia. Across-sectional epidemiological study was carried out from April 2017 to April 2018 to estimate the sero prevalence of bovine and human brucellosis and to assess the associated risk factors of brucellosis in Seka Chokorsa and Shebe Sonbo districts of Jimma zone. Results: The overall prevalence of cattle brucella infection at individual and herd level were 5.9% (95%CI: 4.1%-8.1%) and 26.6 (95%CI: 19.1%-35.3%) based on diagnosis using commercial kits of the competitive enzyme-linked Immunosorbent assay (CELISA) from Brucella abortus antibodies. Univariate logistic regression analysis at individual animal level showed that animals from large herd size and households which had a practice of introduction of new animals in their herds were 3.7 and 2.3 times more likely to be seropositive, respectively. The same scenario has been observed at herd level. Molecularly, five Brucella abortus was identified as the species affecting cattle in the study areas. From the total human serum samples tested only one serum was found to be positive for CFT and thus the prevalence is 0.42 %,( 95%CI: 0.01%-2.38%). From 238 respondents, 90% of them drink raw milk and milk products, and 70% of them also eat raw meat. Furthermore, slaughtering, assisting during delivery and poor management of aborted material is a common practice that favor transmission of the pathogen in the area. Conclusions: In conclusion, the results of this study showed that brucellosis is an important and widely distributed disease in cattle in the study areas. The finding of the infection in human indicates the public health importance of brucellosis in the area. The risk behaviors and practices observed suggests the need to design all-inclusive health programmes, such one-health approach aimed at controlling brucellosis spread in the study area.

scholarly journals Brucellae through the food chain : the role of sheep, goats and springbok (Antidorcus marsupialis) as sources of human infections in Namibia

2011 ◽  
Vol 82 (4) ◽  
Author(s):  
K. Magwedere ◽  
A. Bishi ◽  
G. Tjipura-Zaire ◽  
G. Eberle ◽  
Y. Hemberger ◽  
...  
Keyword(s):  
Goat Milk ◽  
Raw Milk ◽  
Cross Reactivity ◽  
Control Programme ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Serum Samples ◽  
Clinically Normal ◽  
Source Of Infection ◽  
Human Infections

A confirmed case of human brucellosis motivated an investigation into the potential source of infection in Namibia. Since domestic animals are principal sources of Brucella infection in humans, 1692 serum samples were screened from sheep, goats and cattle from 4 presumably at-risk farms and 900 springbok (Antidorcas marsupialis) serum samples from 29 mixed farming units for Brucella antibodies by the Rose-Bengal test (RBT) and positive cases confirmed by complement fixation test (CFT). To assess the prevalence of human brucellosis, 137 abattoir employees were tested for Brucella antibodies using the standard tube agglutination test (STAT) and by enzyme linked immunosorbent assay (ELISA). Cattle and sheep from all 4 farms were negative by RBT and CFT but 2 of the 4 farms (Ba and C) had 26/42 and 12/285 seropositive goats, respectively. Post mortem examination of seropositive goats revealed no gross pathological lesions typical of brucellosis except enlarged mesenteric and iliac lymph nodes seen in a single buck. Culture for brucellae from organs of seropositive animals was negative. None of the wildlife sera tested positive by either RBT or CFT. Interviews revealed that besides the case that prompted the investigation, a family and another person from other farms with confirmed brucellosis shared a common history of consumption of unpasteurised goat milk, home-made goat cheese and coffee with raw milk and prior contact with goats, suggesting goats as the likely source of infection. All 137 abattoir employees tested negative by STAT, but 3 were positive by ELISA. The 3 abattoir workers were clinically normal and lacked historical connections with clinical cases. Although goats are often associated with B. melitensis, these studies could not explicitly implicate this species owing to cross-reactivity with B. abortus, which can also infect goats. Nevertheless, these data reinforce the need for a better National Control Programme for brucellosis in Namibia.

scholarly journals Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay

2012 ◽  
Vol 20 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Neekun Sharma ◽  
Akitoyo Hotta ◽  
Yoshie Yamamoto ◽  
Osamu Fujita ◽  
Akihiko Uda ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Francisella Tularensis ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Novel ◽  
Serum Samples ◽  
Content Type ◽  
Microagglutination Test ◽  
Highly Sensitive ◽  
High Degree

ABSTRACTA novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies againstFrancisella tularensisin humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed againstF. tularensislipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivatedF. tularensis-immunized rabbits, andF. tularensis-infected mice. Antibodies againstF. tularensiswere successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (R2= 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species.

scholarly journals Seroprevalence and Clinical Presentation of Chikungunya Virus Infection among Febrile Outpatients Seeking Health Care in Mzuzu City, Malawi

2021 ◽  
Author(s):  
Flywell Kawonga ◽  
Gerald Misinzo ◽  
Dylo Pemba ◽  
Leonard Mboera ◽  
Isaac Thom Shawa
Keyword(s):  
Chikungunya Virus ◽  
Clinical Presentation ◽  
Age Groups ◽  
Clinical Signs ◽  
Signs And Symptoms ◽  
Viral Disease ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Antibodies

Chikungunya is a mosquito-borne viral disease caused by Chikungunya virus (CHIKV. We conducted this study determine the seroprevalence and clinical presentation of Chikungunya infection among outpatients seeking healthcare in Mzuzu City, Malawi. Blood samples were collected from malaria negative and non-septic febrile outpatients with fevers ≥38 °C, for not more than 5 days. The enzyme- linked immunosorbent assay (ELISA) test was used to detect anti-CHIKV IgM antibodies and its results were used to determine seroprevalence of Chikungunya. A total of 119 serum samples were tested, of these, 73 (61.3%) tested positive for anti-CHIKV IgM antibodies by ELISA. Laboratory requisition forms were used to capture demographic information such as age, sex, clinical signs and symptoms presented by the enrolled patients. Age groups of 1-9, 10- 19, 20- 29, 30- 39, 40- 49, and ≥50 years had 17.8% (n= 13), 12.3 %,( n=9), 15.1%) (n=11), 19.2%; (n=14), 17.8% (n=13) and 17.8% (n=13) proportion of seroprevalence respectively. Most of the CHIKV infected individuals presented with fever (52.05%), joint pain (45.21%) and abdominal pain (42.67%). The presence of anti- CHIKV IgM antibodies suggest the presence of recent CHIKV infection and therefore accurate laboratory assays are highly recommended for CHIKV diagnosis and appropriate management of febrile patients.

scholarly journals Molecular Fine-Specificity Analysis of Antibody Responses to Human Cytomegalovirus and Design of Novel Synthetic-Peptide-Based Serodiagnostic Assays

1999 ◽  
Vol 37 (1) ◽  
pp. 179-188 ◽  
Author(s):  
Astrid E. Greijer ◽  
Jos M. G. van de Crommert ◽  
Servi J. C. Stevens ◽  
Jaap M. Middeldorp
Keyword(s):  
Human Cytomegalovirus ◽  
Immunoglobulin M ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Peptide Epitopes ◽  
Fine Specificity ◽  
Marker Proteins ◽  
Highly Sensitive ◽  
Virion Antigen

To identify single immunodominant marker proteins which can replace complex virion antigen in serodiagnostic assays, we investigated in detail the molecular fine specificity of antibody responses in different individuals with latent or active human cytomegalovirus (HCMV) infection. An overview of the HCMV proteins recognized by human antibodies was obtained by immunoblotting. For selected immunodominant proteins the epitope fine specificity of the antibody response was determined by a peptide-scanning enzyme-linked immunosorbent assay (ELISA). Epitope clusters were synthesized as combination peptides and were used for further serologic analysis of immunoglobulin M (IgM) and IgG reactivities with panels of sera from different groups of patients in comparison to those with cytomegalovirus (CMV) virion antigen. Several serum samples had significantly higher reactivities with peptides than with the CMV virion antigen. However, individual serum samples occasionally recognized diverse peptide epitopes, stressing the importance of using combinations of peptides in serologic assays. From these studies we were able to define a specific combination of peptides derived from pp52 (UL44) and pp150 (UL32) for the specific and highly sensitive early detection of HCMV IgM, whereas a combination of peptides from pp150 (UL32), gB (UL55), and pp28 (UL99) was selected to give optimal and specific reactivity with HCMV IgG. On the basis of the results obtained with these peptide combinations, new, highly specific serodiagnostic assays were constructed. These assays had sensitivities of 98.9 and 96.4% for IgG and IgM, respectively, in comparison with the results obtained with the “gold standard,” the virion antigen-based ELISA. From the results of this study we conclude that specific combinations of highly defined synthetic peptides can replace complex HCMV virion extracts used in current serodiagnostics and may add to further standardization of HCMV serology.

scholarly journals An outbreak of bluetongue virus serotype 9 in Southern Croatia

2011 ◽  
Vol 80 (4) ◽  
pp. 331-336 ◽  
Author(s):  
Eddy Listeš ◽  
Sanja Bosnić ◽  
Miroslav Benić ◽  
Josip Madić ◽  
Željko Cvetnić ◽  
...  
Keyword(s):  
Bluetongue Virus ◽  
Neutralization Test ◽  
Light Trap ◽  
Clinical Signs ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Virus Serotype ◽  
Serum Neutralization Test ◽  
First Time ◽  
Obsoletus Complex

The aim of this study was to provide a description of the first epidemic of bluetongue and the first survey on midges of the genus Culicoides in Croatia. Clinical signs were firstly observed on November 2001 in sheep in Konavle, Dubrovnik – Neretva County. During this epizootic the overall sheep morbidity and mortality were 5.2% (95% confidence interval (c.i.), 4.1-6.6%) and 2.29% (95% c.i., 1.6-3.3%), respectively. After the outbreak, 3,318 serum samples of ruminants from 53 villages of the Dubrovnik – Neretva County were examined for bluetongue virus (BTV) antibodies by competitive enzyme-linked immunosorbent assay (cELISA). In forty nine (92.45%, 95% c.i., 82.11-96.92%) of the 53 villages, animals with antibodies against bluetongue virus were found. In particular, a total of 178 cattle (49.86%, 95% c.i., 44.7-55.0%), 174 sheep (13.72%, 95% c.i., 11.9-15.7%) and 270 goats (15.95%, 95% c.i., 14.3-17.8%) were seropositive. Antibodies to bluetongue virus serotype 9 were detected in 212 positive sera by serum neutralization test. The percentage of positive animals decreased (P > 0.05) from the east to the west suggesting a possible east westward spreading of BTV infection. Fourteen light-trap midge collections from seven different sites were examined. Of the 4872 Culicoides spp. collected, 4,492 (92%, 95% c.i., 91.4-92.9%) of them belonged to the species of Obsoletus complex. This study showed for the first time that a pathogenic strain of BTV-9, probably from Montenegro, entered Croatia causing disease and death in local sheep and that C. obsoletus and C. scoticus were likely the major vectors of infection.

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 Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Harisankar Singha ◽  
Praveen Malik ◽  
Sachin K. Goyal ◽  
Sandip K. Khurana ◽  
Chiranjay Mukhopadhyay ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Expression System ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Efficacy ◽  
Diagnostic Potential ◽  
Prokaryotic Expression System ◽  
Control Serum

Objective. To express truncated TssB protein ofBurkholderia malleiand to evaluate its diagnostic efficacy for serological detection of glanders among equines.Materials and Methods. In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences ofB. malleiTssB protein—a type 6 secretory effector protein—were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (n=49), negative (n=30), and field serum samples (n=1811). Cross-reactivity of the assay was assessed with equine disease control serum and human melioidosis positive serum.Results. In comparison to CFT, diagnostic sensitivity and specificity of ELISA were 99.7% and 100%, respectively.Conclusions. The indirect ELISA method using the truncated TssB offered safer and more rapid and efficient means of serodiagnosis of glanders in equines. These data highlight the use of TssB as potential diagnostic antigen for serological diagnosis of glanders.

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