2012, Number 3
<< Back Next >>
Rev Cubana Med Trop 2012; 64 (3)
Increase of analytical sensitivity of FasciDIG® system for the diagnosis of Fasciola hepatica
Marcet SR, Figueredo PM, Núñez FFA, Rojas RL, Sarracent PJ
Language: Spanish
References: 18
Page: 335-341
PDF size: 72.31 Kb.
ABSTRACT
Introduction: fasciolosis is an endemic disease in cattle in Cuba and there is an increase in the number of reported human cases in recent years. The copro-parasitological diagnosis of fasciolosis has low sensitivity and is hard-working; for that reason, it is important to use immunoenzymatic methods mainly those that can detect this parasite antigens in the feces. A system for antigen detection called FasciDIG®, with a reported sensitivity of 10 ng/mL has been developed in "Pedro Kouri" Institute of Tropical Medicine.
Objective: to increase the sensitivity of FasciDIG ® through some modifications to this diagnostic method.
Methods: two foul dilutions (concentrations of antigen 1 000 ng/mL- 1.95ng/mL in H2O Tween-20) were evaluated in a simulated system using FasciDIG® and modified FasciDIG. The FasciDiG® was modified using the secondary antibody obtained from rabbit against excretory-secretory antigens of Fasciola hepatica combined with biotin and then adding commercial conjugated extravidine peroxidase. Feces were collected from the rectum of 96 animals for slaughter and were evaluated by both methods, FasciDig® and modified FasciDig. Kappa index was calculated between both assays.
Results: the detection limit for the FasciDIG® was 3.9 ng/mL whereas the modified FasciDIG detected up to 1.95 ng/mL. The agreement index calculated between the two tests was 0.6238 corresponding to an index of substantive or good agreement.
Conclusions: the modified method is more sensitive than FasciDIG® and it can supplement the diagnosis of fasciolosis. The number of analyzed samples should be increased and the sensitivity and specificity should also be determined using the serial conic-cup sedimentation technique as the gold standard.
REFERENCES
Mas-Coma S. Human fascioliasis. En: Cotruvo JA, Dufour A, Rees G, Bartram J, Carr R, Cliver DO, et al., editors. Waterborne zoonoses: Identification, causes and control. London, UK: World Health Organization (WHO); 2004. p. 305-22.
Mas-Coma S. Epidemiology of fascioliasis in human endemic areas. J Helminthol. 2005;79:207-16.
Karahocagil MK, Akdeniz H, Sunnetcioglu M, Cicek M, Mete R, Akman N, et al. A familial outbreak of fascioliasis in Eastern Anatolia: a report with review of literature. Acta Trop. 2011;118:177-83.
Maguire JH. Schistosomiasis. En: Mandel GL, Bennett JE , Dolin R, editors. Trematodes and other flukes. New York: Churchill Livingstone; 2005: 3276-85.
Charlier J, Meulemeester L, Claerebout E, Williams D, Vercruysse J. Qualitative and quantitative evaluation of coprological and serological techniques for the diagnosis of fasciolosis in cattle. Vet Parasitol 2008;153:44-51.
Spithill TW SP, Sexton LS, Bozas E, Morrison CA, Creaney J, Parsons JC. Devellopment of vaccine against of Fasciola hepatica. In: JPD, editors. Fasciolosis. Ireland: Wallingford: CAB International; 1999. p. 377-410.
Rojas L, Vazquez A, Domenech I, Robertson LJ. Fascioliasis: can Cuba conquer this emerging parasitosis? Trends Parasitol. 2010;26:26-34.
Espino AM, Marcet R, Finlay CM. Fasciola hepatica: detection of antigenemia and coproantigens in experimentally infected rats. Exp Parasitol. 1997;85:117-20.
Espino AM, Millan JC, Finlay CM. Detection of antibodies and circulating excretory-secretory antigens for assessing cure in patients with fascioliasis. Trans R Soc Trop Med Hyg. 1992;86:649.
Espino AM, Marcet R, Finlay CM. Detection of circulating excretory secretory antigens in human fascioliasis by sandwich enzyme-linked immunosorbent assay. J Clin Microbiol. 1990;28:2637-40.
Mezo M, Gonzalez-Warleta M, Carro C, Ubeira FM. An ultrasensitive capture ELISA for detection of Fasciola hepatica coproantigens in sheep and cattle using a new monoclonal antibody (MM3). J Parasitol. 2004;90:845-52.
Espino AM, Diaz A, Perez A, Finlay CM. Dynamics of antigenemia and coproantigens during a human Fasciola hepatica outbreak. J Clin Microbiol. 1998;36:2723-6.
Espino AM. Inmunodianóstico de la Fasciolosis humana y su aplicación en brotes epidémicos [Doctoral thesis]. La Habana: Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí"; 1997. p. 65.
Gupta A, Dixit AK, Dixit P, Mahajan C, Sharma RL. Evaluation of dipstick-ELISA using 28 kDa Fasciola gigantica cathepsin l cysteine proteinase (FgCL3) for serodiagnosis of fasciolosis in naturally infected goats. Vet Parasitol. 2011;176:165-9.
Rigby AS. Statistical methods in epidemiology. v. Towards an understanding of the kappa coefficient. Disabil Rehabil. 2000;22:339-44.
Crowther JR. ELISA Theory and Practice. Totowa, New Jersey: Human Press; 1995. 1-219 p.
Ubeira FM, Muino L, Valero MA, Periago MV, Perez-Crespo I, Mezo M, et al. MM3-ELISA detection of Fasciola hepatica coproantigens in preserved human stool samples. Am J Trop Med Hyg. 2009;81:156-62.
Espinoza JR, Maco V, Marcos L, Saez S, Neyra V, Terashima A, et al. Evaluation of Fas2-ELISA for the serological detection of Fasciola hepatica infection in humans. Am J Trop Med Hyg. 2007;76:977-82.