Comparison between in-house indirect ELISA and Dot-ELISA for the diagnosis of Fasciola gigantica in cattle

Yamchi, Jafar Arjmand; Hajipour, Nasser; Froushani, Seyyed Meysam Abtahi; Keighobadi, Mojtaba

doi:10.1007/s12639-015-0699-4

Cited by 4 publications

(1 citation statement)

References 22 publications

(20 reference statements)

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“…The SDS-PAGE analysis of Fg-E/S antigens in the present study showed that many bands have appeared, and the most prominent of which were of 15, 17, 28, 40, 60 and 84 kDa. Some bands (40, 60, and 84) were shown to be derived from the worm tegument (Viyanant et al, 1997), and they are the major recognition targets in ESP-based immunodiagnosis, while the 15,17 and 28 kDa species appeared to be released from the cells lining the gut (Anuracpreeda et al, 2006;Arjmand Yamchi et al, 2016). (Awad et al, 2009) showed that the Fg-ES antigen revealed seven protein bands with molecular weights of 15, 28, 31.6, 32.9, 39.4, 83.3 and 101.7 kDa.…”

Section: Discussionmentioning

confidence: 99%

Serological Immunodetection of Fasciola gigantica Excretory/ Secretory Antigens in Naturally Infected Cattle and Human

Khalil

Ibrahim

2023

Egyptian Academic Journal of Biological Sciences, B. Zoology

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The traditional diagnostic method of fascioliasis was based on the identification of the eggs in the stool, but this is not a reliable way as it has many restrictions. The present work was evaluated to immunodetect the Excretory/ Secretory antigens of Fasciola gigantica by sandwich Elisa in naturally infected cattle's sera and by dot blot in naturally infected human sera and compare these methods and the traditional methods. In this study, fresh adult Fasciola gigantica worms were collected to extract crude excretory/secretory (E/S) antigens. E/S was used to immunize rabbits and mice to raise polyclonal antibodies. The IgG fraction of rabbit and mouse anti-Fasciola antibodies was purified. The protein content of anti-Fasciola IgG antibody, was 8.8 and 5.2mg/ml, respectively. Sandwich ELISA was performed to detect Fasciola antigens in serum samples collected from 248 cattle. Also, a Dot blot was performed to detect Fasciola antigens in 38 human sera. Results showed that after parasitological stool examination, 26 human samples were positive and 12 human sera were healthy. The redbrown color appeared with all infected samples only. This technique is cheap, and saves time; multiple samples can make at the same time and not require expensive laboratory equipment. In conclusion, the sandwich ELISA and Dot blot assays were more reliable tools for early serodiagnosis of fasciolosis than traditional methods. Significance Statement: Different antigenic fractions of Fasciola have been used for serological diagnosis of human fasciolosis.

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Section: Discussionmentioning

confidence: 99%

Serological Immunodetection of Fasciola gigantica Excretory/ Secretory Antigens in Naturally Infected Cattle and Human

Khalil

Ibrahim

2023

Egyptian Academic Journal of Biological Sciences, B. Zoology

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Prevalence of bovine fascioliasis in a new-emerging focus of human fascioliasis in BoyerAhmad district, southwest of Iran

Zaraei

Arefkhah

Moshfe

et al. 2019

Comparative Immunology, Microbiology and Infectious Diseases

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Sensitivity and specificity for African horse sickness antibodies detection using monovalent and polyvalent vaccine antigen-based dot blotting

et al. 2022

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Background and Aim: The immune responses of animals infected with African horse sickness (AHS) virus are determined by enzyme-linked immunosorbent assay (ELISA), complement fixation, and virus neutralization test. During the outbreaks of AHS in Thailand, the immune response after vaccination has been monitored using commercial test kits such as blocking ELISA, which are expensive imported products unavailable commercially in Thailand. This study aimed to assess the sensitivity and specificity of anti-AHS virus antibodies using dot blotting based on monovalent and polyvalent strains of live attenuated AHS vaccine. Materials and Methods: A total of 186 horse sera, namely, 93 AHS-unvaccinated samples and 93 AHS-vaccinated samples, were used in this study. All sera underwent antibodies detection using commercial blocking ELISA and in-house dot blotting based on monovalent and polyvalent strains of live attenuated AHS vaccine. The numbers of true positive, false positive, true negative, and false negative results in the dot blotting were compared with those in blocking ELISA and the sensitivity and specificity of dot blotting were assessed. Results: For the monovalent antigen, there were 78, 19, 74, and 15 true positive, false positive, true negative, and false negative results, respectively, while for the polyvalent antigen, the corresponding numbers were 84, 34, 58, and 9. Meanwhile, the diagnostic sensitivity and specificity for monovalent antigen were 83.87% and 79.57%, respectively, but 90.32% and 62.37% for polyvalent antigen. Conclusion: Dot blotting for AHS antibodies detection using vaccine antigen showed high sensitivity and rather a high specificity compared with the findings with the commercial ELISA test kit. In countries where commercial ELISA test kits are not available and when the size of a serum sample is small, dot blotting could become a good alternative test given its advantages, including its simplicity, rapidity, and convenience. To the best of our knowledge, these findings are the first report on the use of dot blotting for detecting AHS antibodies in horses. In conclusion, monovalent antigen-based dot blotting could be used as a reliable alternative serodiagnostic test for monitoring AHS humoral immune response, especially in vaccinated horses.

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Comparison between in-house indirect ELISA and Dot-ELISA for the diagnosis of Fasciola gigantica in cattle

Cited by 4 publications

References 22 publications

Serological Immunodetection of Fasciola gigantica Excretory/ Secretory Antigens in Naturally Infected Cattle and Human

Serological Immunodetection of Fasciola gigantica Excretory/ Secretory Antigens in Naturally Infected Cattle and Human

Prevalence of bovine fascioliasis in a new-emerging focus of human fascioliasis in BoyerAhmad district, southwest of Iran

Sensitivity and specificity for African horse sickness antibodies detection using monovalent and polyvalent vaccine antigen-based dot blotting

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