scholarly journals Evaluation of a bovine pregnancy-associated glycoprotein enzyme-linked immunosorbent assay kit for serological diagnosis of pregnancy in sheep

2015 ◽  
Vol 46 (2) ◽  
pp. 362-367 ◽  
Author(s):  
Monique Tomazele Rovani ◽  
Alfredo Skrebsky Cezar ◽  
Melânia Lazzari Rigo ◽  
Bernardo Garziera Gasperin ◽  
Janduí Escarião da Nóbrega Júnior ◽  
...  

ABSTRACT: Pregnancy diagnosis is an important tool for farm management. Ultrasonography is the main technique used for pregnancy diagnosis in ewes. As an alternative, radioimmunoassay (RIA) allows accurate and early detection of pregnancy-associated glycoproteins (PAGs) in sheep blood. However, radioactive-based techniques, as RIA, have been increasingly inadvisable due to environmental risk. Homology between ovine and bovine PAGs is high, and ELISA kits used for PAGs detection in cattle are safer than RIA. Thus, this study aimed to evaluate the feasibility of PAGs detection for pregnancy diagnosis in sheep serum samples using an ELISA kit produced for cattle. The sensitivity and specificity of the ELISA kit were 93.5% and 98.9%, respectively, whereas positive and negative predictive values were 99.0% and 93.1%, respectively, in comparison to ultrasonography diagnostic (control). PAGs reached consistently detectable concentrations in ovine serum around 33 days after mating. Accuracy of the ELISA test was 96.1% from 33 days of pregnancy until lambing. After parturition, PAGs were still detectable seven days post-lambing. However, from 21 days post-parturition, PAGs from the previous pregnancy were no longer detected in serum samples. In conclusion, the bovine ELISA kit can accurately detect pregnancy in sheep 33 days following mating, while PAGs levels from the previous gestation are no longer detected from 21 days post-partum. The evaluated ELISA test is a reliable tool for pregnancy diagnosis in sheep at random stages and as a complementary exam at early gestation.

2012 ◽  
Vol 45 (4) ◽  
pp. 510-513 ◽  
Author(s):  
Teiliane Rodrigues Carneiro ◽  
Marta Cristhiany Cunha Pinheiro ◽  
Sara Menezes de Oliveira ◽  
Ana Lúcia de Paula Hanemann ◽  
José Ajax Nogueira Queiroz ◽  
...  

INTRODUCTION: The laboratory diagnosis of schistosomiasis is based mainly on the detection of parasite eggs in stool samples through the Kato-Katz (KK) technique, reading one slide by test. However, a widely known limitation of parasitological methods is reduced sensitivity, particularly in low endemic areas. METHODS: To increase sensitivity, we conducted further slide readings from the same stool sample using the parasitological method associated with a serological test. We used the KK method (three slides) and the IgG anti-Schistosoma mansoni-enzyme-linked immunosorbent assay (ELISA) technique to diagnose schistosomiasis in low endemic areas in the Brazilian State of Ceará. Fecal samples and sera from 250 individuals were analyzed. RESULTS: Sixteen percent and 47.2% of samples were positive in parasitological tests and serological tests, respectively. Parasitological methods showed that 32 (80%) individuals tested positive on the first slide, 6 (15%) on the second slide, and 2 (5%) on the third. The performance of the ELISA test in the diagnosis, using the KK method as diagnostic reference, showed a negative predictive value of 100%, with specificity and positive predictive values of 62.8% and 33.9%, respectively. CONCLUSIONS: In this study, the increase from one to three slides analyzed per sample using the KK technique was shown to be a useful procedure for increasing the diagnostic sensitivity of this technique.


2013 ◽  
Vol 21 (1) ◽  
pp. 96-106 ◽  
Author(s):  
Lourena E. Costa ◽  
Mayara I. S. Lima ◽  
Miguel A. Chávez-Fumagalli ◽  
Daniel Menezes-Souza ◽  
Vivian T. Martins ◽  
...  

ABSTRACTVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy andTrypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected withLeishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n= 31) compared to those from vaccinated dogs (n= 21), experimentally infected dogs with cross-reactive parasites (n= 23), and healthy controls (n= 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no cross-reactivity withT. cruzi- orEhrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes ofL. infantumantigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs.


1996 ◽  
Vol 32 (5) ◽  
pp. 397-400
Author(s):  
SD Babyak ◽  
MG Groves ◽  
DS Dimski ◽  
J Taboada

An enzyme-linked immunosorbent assay (ELISA) test kit for the detection of feline leukemia virus (FeLV) antigen in saliva was evaluated in 150 cats. Saliva and blood samples from all cats were tested for FeLV using the saliva ELISA kit and a plasma ELISA kit, respectively. These results were compared with indirect immunofluorescent antibody (IFA) testing of blood smears also obtained from each cat. The proportion of cats that tested positive were 10%, 7%, and 8% for each test, respectively. Using the IFA test as the gold standard, the saliva FeLV test had a sensitivity of 91.7% and specificity of 97.1%, while the plasma ELISA test had a sensitivity of 91.7% and specificity of 100%.


Author(s):  
Ayse Kilic ◽  
Hakan Kalender

Q fever is a zoonotic disease that occurs worldwide and is caused by the obligate intracellular bacterium Coxiella burnetii. Infected animals are usually asymptomatic, but infection can cause abortion and stillbirth in ruminants. The main purpose of this study was to evaluate prevalance of Coxiella burnetii infection in aborted and nonaborted sheep serum samples in Eastern Anatolia region by using enzyme-linked immunosorbent assay (ELISA). The determine of prevalance in sheep flocks from four provinces (Elazig, Malatya, Tunceli, Bitlis) and tested for anti-C.burnetii antibody detection, by means of Chekit Q fever Elisa kit. 350 serum samples obtained from flocks belonging aborted sheep showed that a total of 56 (16%) were detected seropositivity, whereas 171 serum samples obtained from nonaborted sheep flocks in 13 of the 171 (7.60%) for C.burnetii in seropositivity were observed. Coxiellosis should be considered an important cause of sheep with abortion history and nonaborted in Elazig and neighboring provinces.


Author(s):  
Daniel Muasya ◽  
George Karuoya Gitau ◽  
Thaiyah Gitau Andrew ◽  
Daniel Waweru Gakuya ◽  
John Vanleeuwen ◽  
...  

Bovine tuberculosis (BTB) is an important zoonotic disease which has remained persistent in many areas of the World. Control and eradication has proved problematic due to the challenges in effective screening and diagnosis. Limited information on the status of BTB in Kenya presents a need to investigate its occurrence in cattle population. The study was carried out using an indirect antibody Enzyme Linked Immunosorbent Assay (ELISA-IDEXX-USA) to assess the agreement with Comparative Intradermal Tuberculin Test (CITT) test results between August and December 2013 in Laikipia County. The study also determined the apparent prevalence of cattle BTB antibodies in Laikipia County. A total of 276 bovine serum samples were tested using MPB70 and MPB83 recombinant proteins as capture antigens in the ELISA kit. Data were recorded in Microsoft Excel and exported to SPSS 16.0 for analysis. Apparent prevalence was calculated as a proportion at 95% CI, Kappa statistics computed, and test of significance assessed by Chi-square and Fisher exact test.  The results showed an individual animal apparent prevalence of 3.9% (11/276) and a herd prevalence of 58.3% (7/12). There was no significant association between BTB infection and both animal level factors, breed and sex. Kappa agreement test between ELISA test and TST showed a good agreement at K= 0.65. This study reported the prevalence of BTB in cattle for the first time in Laikipia County Kenya using the IDEXX ELISA. The ELISA test was comparable to the tuberculin test which is used as the gold standard for screening TB on live animals. The two tests can be used alongside for series or parallel interpretation to achieve desirable diagnostic sensitivity and specificity in disease control programs.


Author(s):  
Flywell Kawonga ◽  
Gerald Misinzo ◽  
Dylo Pemba ◽  
Leonard Mboera ◽  
Isaac Thom Shawa

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.


2019 ◽  
Vol 2 (2) ◽  
pp. 64
Author(s):  
Ni Ketut Suwiti ◽  
Luh Gde Surya Heryani ◽  
Desak Nyoman Dewi Indira Laksmi ◽  
Ni Nyoman Werdi Susari ◽  
I Nengah Kerta Besung ◽  
...  

The aim of this research was to detect identify levels of Bovine Immunoglobulin E (BoIg.E), can be used as an indicator of response immune in bali cattle.  Eighty serum samples were collected from Nusa Penida and Bangli region. Bovine Ig.E levels was measured using a commercial Enzyme Linked Immunosorbent Assay (ELISA) Kit. The data were analysis based on differences of farming characteristics andgeographic. The result of research that, of BoIg.E level of bali cattle kept in Bangli (34.16258 ?g/ml), was higher than Nusa Penida (22.26047 ?g/ml). We conclude that there was a significant effect of differences of farming characteristics and geographic conditions.


2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Mwemezi L. Kabululu ◽  
Maria V. Johansen ◽  
James E. D. Mlangwa ◽  
Ernatus M. Mkupasi ◽  
Uffe C. Braae ◽  
...  

Abstract Background Taenia solium is a zoonotic parasite responsible for neurocysticercosis—a major cause of late-onset acquired epilepsy in humans. Lack of affordable, specific and sensitive diagnostic tools hampers control of the parasite. This study assessed the performance of an antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) in the diagnosis of viable T. solium cysticercosis in naturally infected slaughter-age pigs in an endemic area in Tanzania. Methods A total of 350 pigs were bled before they were slaughtered and their carcases examined. Serum was analyzed for circulating antigens by using a monoclonal antibody-based B158/B60 Ag-ELISA. Each carcase was examined for the presence of Taenia hydatigena cysticerci and half carcase musculature together with the whole brain, head muscles, tongue, heart and diaphragm were sliced with fine cuts (< 0.5 cm) to reveal and enumerate T. solium cysticerci. Half carcase dissection can detect at least 84% of infected pigs. Prevalence and their 95% confidence intervals (CI) were calculated in Stata 12. Sensitivity, specificity, predictive values and likelihood ratios were determined. Results Twenty–nine pigs (8.3%, 95% CI: 5.6–11.7%) had viable T. solium cysticerci while 11 pigs had T. hydatigena cysticerci (3.1%, 95% CI: 1.6–5.5%). No co-infection was observed. Sixty-eight pigs (19.4%, 95% CI: 15.4–20%) tested positive on Ag-ELISA; of these, 24 had T. solium cysticerci and 7 had T. hydatigena cysticerci. Sensitivity and specificity were determined to be 82.7% and 86.3%, respectively. Positive and negative predictive values were 35.2% and 98.2%, respectively. Likelihood ratios for positive and negative Ag-ELISA test results were 6.0 and 0.2, respectively. There was a significant positive correlation between the titre of circulating antigens and intensity of T. solium cysticerci (r(348) = 0.63, P < 0.001). Conclusions The Ag-ELISA test characteristics reported in this study indicate that the test is more reliable in ruling out T. solium cysticercosis in pigs, than in confirming it. Hence, a negative result will almost certainly indicate that a pig has no infection, but a positive result should always be interpreted with caution. Estimates of T. solium prevalence based on Ag-ELISA results should, therefore, be adjusted for test performance characteristics and occurrence of T. hydatigena.


Author(s):  
Naser Nazari ◽  
Tooran Nayeri ◽  
Farkhondeh Hazrati

Background and Aims: Echinococcus granulosus (E. granulosus) is a cestode parasite that causes cystic hydatid disease in humans worldwide. Iran is one of the endemic regions for infection that indicate the importance and presence of infection in this country. Therefore, the current research aimed to characterize the seroprevalence of human cystic echinococcosis in Sanandaj city, Kurdistan province, western Iran. Materials and methods: Totally, 500 serum samples were collected from patients referred to different health centers in Sanandaj city using cluster sampling in 2018-2019. All the sera were examined using the enzyme-linked immunosorbent assay test. Results: The seroprevalence of human hydatidosis was reported at 2.2% by ELISA test in Sanandaj city. This rate was 9 (1.9%) in women and 2 (0.4) in men. The age group of 20-30 years old had the highest positivity rate (1.0%). Also, the subjects that consumed home slaughtered meat had the highest infection rate at 4 (0.8%). There was no significant difference regarding factors studied such as sex, education, residence, consumed water, keeping a dog, and the seropositivity. Conclusions: Seroprevalence of human cystic echinococcosis in Sanandaj city is lower than the general prevalence in Iran. Our research team hopes to provide accurate data on the prevalence of hydatidosis in Sanandaj encourage more extensive research to help prevent this parasite in Iran and worldwide.


2004 ◽  
Vol 11 (4) ◽  
pp. 775-779 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Hisham Ismail ◽  
Gellan G. Ibrahim ◽  
Mohamed Abdel-Raouf ◽  
Ahmed M. El-Waseef ◽  
...  

ABSTRACT Recently, noninvasive diagnostic tests for Helicobacter pylori infection have gained in significance. We have developed a sensitive and specific noninvasive immunoassay based on the detection of an H. pylori circulating antigen (HpCA) in sera from H. pylori-infected individuals. Monospecific antibody and Western blot analyses were used to demonstrate the presence of the target antigen in H. pylori cell lysate and serum samples. A novel enzyme-linked immunosorbent assay (ELISA) was developed for the detection of HpCA in serum. Endoscopic biopsy specimens from the gastric antra of 221 individuals (143 males and 78 females) with dyspeptic symptoms were evaluated for H. pylori infection, with culture used as a “gold standard” for diagnosis. The target H. pylori antigen was identified at 58 kDa. HpCA has been detected by ELISA with high degrees of sensitivity, specificity, and efficiency (>90%), and ELISA results show no significant difference (P > 0.05) from results of H. pylori culture of gastric biopsy specimens. The test's positive and negative predictive values were also high (95 and 86%, respectively). In conclusion, a sensitive and specific immunoassay was developed for the detection of HpCA in human serum. This test can be applied for noninvasive laboratory and field diagnoses of H. pylori infection.


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