Kiran AFSHAN, Imtiaz AHMAD, Maria KOMAL, Sabika FIRASAT, Imtiaz Ahmad KHAN, Mazahr QAYYUM

Diagnostic Efficacy of Copro-ELISA for Detection of Fasciolosis in Cattle and Buff aloes in Punjab Province, Pakistan

Fasciolosis is a food borne zoonotic trematode disease that causes liver damage in ruminants and humans. The information recorded on the prepatent diagnostic methods is limited with low sensitivity. The study aimed to investigate the diagnostic efficacy of coproantigens by using in house developed indirect enzyme linked immunosorbent assay (ELISA). Adult helminths were collected from the bile duct of buff aloes for the extraction of excretory secretory (ES) and somatic (SA) antigens. The polyclonal antibodies were produced by immunization of rabbits with SA and ES antigens of Fasciola. Kappa value of developed ELISA was calculated to check the diagnostic performance of the test. The mean absorbance values at diff erent concentrations of coproantigens were significantly diff erent (P≤0.001) from controls. However, the diff erence was not significant among the concentrations of coproantigen. A positive linear relationship was observed to the concentration of antigens used in fecal supernatant and the absorbance values. The sensitivity and specificity of diagnostic test with ES-polyclonal antibodies were 100% (95% CI: 89.42%-100.00%) and 76.19% (95% CI: 52.83.30%-91.78%), respectively. Kappa value revealed that the strength of agreement is almost substantial. The SA-polyclonal antibodies showed the specificity and sensitivity of diagnostic test were 100% (95% CI: 89.42%-100.00%) and 90.00% (95% CI: 68.30%-98.77%), respectively. However, Kappa value of the test revealed that the strength of agreement is perfect. The result provides information that will add sensitive diagnostic methods for the detection of fasciolosis.

Pakistan’ın Pencap Eyaletinde Sığır ve Mandalarda Fasciolozis’in Tespitinde Copro-ELISA’nın Tanısal Etkinliği

Fasciolozis, geviş getiren hayvanlarda ve insanlarda karaciğer hasarına neden olan gıda kaynaklı zoonotik bir trematod hastalığıdır. Prepatent tanı yöntemleri ile elde edilen diagnostik bilgiler düşük hassasiyetli ve sınırlıdır. Bu çalışma, in house geliştirilen indirekt Enzyme Linked Immunosorbent Assay (ELISA) kullanılarak koproantijenlerin tanısal etkinliğinin araştırılmasını amaçlamıştır. Ekskretuar sekretuar (ES) ve somatik (SA) antijenlerin ekstraksiyonu için kullanılacak erişkin helmintler bufaloların safra kanallarından toplandı. Fasciola’nın SA ve ES antijenleri ile tavşanların immunizasyonunu takiben poliklonal antikorlar üretildi. Geliştirilen ELISA testinin tanısal performansını kontrol etmek için Kappa değeri hesaplandı. Koproantijenlerin farklı konsantrasyonlardaki ortalama absorbans değerleri, kontrollerden önemli ölçüde farklı saptandı (P≤0.001). Fakat, koproantijen konsantrasyonları arasındaki fark önemli değildi. Fekal süpernatanda kullanılan antijenlerin konsantrasyonu ile absorbans değerleri arasında pozitif linear bir ilişki gözlemlendi. ES-poliklonal antikorlarla yapılan diagnostik testin duyarlılığı %100 (%95 CI: %89.42-100.00) ve özgüllüğü %76.19 (%95 CI: %52.83-91.78) saptandı. Kappa değeri, uyum gücünün neredeyse önemli derecede olduğunu ortaya koydu. SA-poliklonal antikorlarlarla yapılan diagnostik testin özgüllüğü %100 (%95 CI: %89.42-100.00) ve duyarlılığı %90.00 (%95 CU: %68.30-98.77) saptandı. Ancak, testin Kappa değeri, uyum gücünün mükemmel olduğunu ortaya koydu. Bulgular, fasciolozisin tespiti için hassas tanı yöntemlerine ek bilgi sağlayacak niteliktedir.

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