Comparison of Dot-ELISA and ELISA Techniques for Detection of Fasciola hepatica in Sheep Using Excretory-Secretory Antigens

Abstract

Fascioliasis is one of the most important zoonosis with worldwide distribution, which causes great losses in livestock production. Serological methods are used for early detection of disease. The current investigation undertook to challenge dot-ELISA and ELISA techniques in diagnosing of Fasciola hepatica in sheep using excretory-secretory (E/S) antigen to select the most appropriate method. The serum samples (n=281) were collected from 50 cases of fascioliasis, 81 animals with parasitic diseases other than fascioliasis, and 150 healthy control cases. The E/S antigen used was obtained from infected livers (F. hepatica) of sheep slaughtered in Hamedan industrial abattoir (Iran). The result of dot-ELISA showed 100% sensitivity and 92.2% specificity. Also, positive and negative predictive values, and accuracy of assay were 73.52%, 100% and 93.59%, respectively. In the case of ELISA, the results were as follows: 98% sensitivity, 100% specificity, 100% positive predictive values, 99.56% negative predictive values, and 99.64% accuracy of assay. In both techniques, cross reaction with fascioliasis was not observed. In conclusion, although these two tests had very similar results, dot-ELISA was more acceptable with respect to its higher sensitivity, simplicity in practice and cheaper than ELISA. Therefore, dot-ELISA can be recommended as a routine test in paraclinical laboratories and epidemiological studies.

Keywords

• Fasciola hepatica, excretory-secretory

Koyunlarda Fasciola hepatica Enfeksiyonunun Tespitinde Ekskresyon / Sekresyon Antijenleri Kullanılarak Dot-ELISA ve ELISA Tekniklerinin Karşılaştırılması

Abstract

Fasiolazis, dünya çapında hayvancılıkta büyük kayıplara neden olan önemli zoonozlardan biridir. Hastalığın erken tanısı için serolojik metotlar kullanılmaktadır. Bu araştırmada, koyunlarda Fasciola hepatica enfeksiyonlarının ekskresyon/sekresyon antijenleri kullanılarak tanısında Dot-ELISA ve ELISA tekniklerinden hangisinin en uygun olduğu karşılaştırılmıştır. Serum numuneleri (n=281) 50 adet fascioliasis olgusundan, 81 adet fasiolasis dışındaki paraziter hastalık olgusundan ve 150 adet sağlıklı kontrol olgularından toplanmıştır. Kullanılan E/S antijeni, Hamedan mezbahasında (İran) kesilen enfekte koyun karaciğerinden (F. hepatica) temin edilmiştir. Dot ELISA sonuçları %100 duyarlılık ve %92,2 belirleyicilik göstermiştir. Ayrıca, pozitif, negatif tahmini değerler ve testin doğruluğu sırasıyla %73,52, %100, %93,59 bulunmuştur. ELISA metodunda sonuçlar, %98 duyarlılık, %100 belirleyicilik, %100 pozitif tahmini sonuç, %99,56 negatif tahmini sonuç ve %99,64 testin doğruluğu olarak belirlenmiştir. Her iki teknikte de fasiyolazis ile çapraz reaksiyon gözlenmemiştir. Sonuç olarak bu iki testin benzer sonuçları olmasına rağmen, Dot ELISA metodu, ELISA metoduna göre daha yüksek duyarlılığa sahip olması, uygulamadaki basitliği ve ucuz olması açısından daha kabul edilebilirdir. Bu nedenle Dot-ELISA, paraklinik laboratuvarlarında ve epidemiyolojik çalışmalarda rutin bir test olarak tavsiye edilebilir.

Keywords

• Fasciola sekresyon antijenleri, Dot-ELISA, ELISA, koyun

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