Improved serodiagnosis of hydatid cyst disease using gold nanoparticle labeled antigen B in naturally infected sheep.

  • Zahra Jahani Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
  • Behnam Meshgi Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
  • Masomeh Rajabi-Bzl Dept. of Medical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
  • Fatemeh Jalousian Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
  • Sajad Hasheminasab Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
Keywords: Antigen B, Diagnosis, Hydatid cyst, Nanoparticle


Background: Cystic echinococcosis caused by the metacestode of Echinococcus granulosus is a major problem in both humans and domestic animals health. There-fore, a standardized and approachable diagnostic tool (rapid tests) for the serodiag-nosis of cystic echinococcosis (CE) is still needed.Methods: In the present work, antigen B labeled with gold nanoparticles was used to detect antibodies against hydatid cyst disease. The prepared antigen B was ana-lyzed by SDS-PAGE. Tetra chloroauric acid (HAuCl4) was used to produce colloi-dal gold and antigen B labeled by gold nanoparticles, then it was tested by using rabbits antisera and sera from naturally infected sheep. The labeled antigen B was evaluated using Dot-immunogold staining (Dot-IGS) method.Results: Electrophoretic pattern of hydatid cyst fluid showed the quality of bands in the condensed fluid is better than crude fluid. SDS-PAGE analysis cyst fluid and antigen B revealed three specific protein bands that were detected at molecular weights of 24, 30 and 40 kDa that all are the subunits of antigen B. Evaluation of antigen B labeled by gold nanoparticles by using Dot-IGS technique showed 1/1 and 1/50 dilutions in comparison with another has the best immunoreaction. In this method, nanoparticles produced a typical purple color, when they binded to the strip at the site of immunoreaction.Conclusion: Therefore, using gold nanoparticles is a good candidate for detection of helminthiasis, also as selective tools of early detection, simple and cost-effective, regardless of specific skills and equipment with optimal durability.


Kittelberger R, Reichel MP, Jenner J, Heath DD, Lightowlers MW, Moro P, Ibrahem MM, Craig PS, Keefe JS. Evaluation of three en-zyme–linked immunosorbent assays for the de-tection of serum antibodies in sheep infected with Echinococcus granulosus. Vet Parasitol. 2002;110:57-76.

Zhang W, Li J, McManus DP. Concepts in immunology and diagnosis of hydatid disease. Clin Microbiol Rev. 2003;16:18-36.

Biava MF, Doa A, Fortier B. Laboratory diag-nosis of cystic hydatid disease. World J Surg. 2000;5(14):22-27.

Eckert J, Deplazes P. Biological, epidemiologi-cal and clinical aspects of echinococcosis, a zo-onosis of increasing concern. Clin Microbiol Rev. 2004;17:107-135.

Grimm F, Maly FE, Lu J, Llano R. Analysis of specific immunoglobulin G subclass antibodies for serological diagnosis of Echinococcosis by a standard enzyme-linked immunosorbent as-say. Clin Diag Lab Immunol. 1998;5:613-616.

Virginio VG, Hernandez A, Rott MB, Monteiro KM, Zandonai AF, Neito A, Zaha A, Ferreira HB. A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease. Clin Exp Immunol. 2003; 132:75-82.

Faulk WP, Taylor GM. An immunocolloidal method for the ekectron microscope. Immu-nochem. 1971;8:1081.

Gupta R, Talwar GP, Gupta SK. Rapid anti-body capture assay for detection of Group A Streptococci using monoclonal antibody and colloidal gold-monospecific polyvalent anti-body conjugate. J I mmunol. 1992;13:441.

Wang JY, Shi F, Yang YT, Gao CH, Bao YF, Tang LH. Establishment and evaluation of col-loidal gold labeled immunochromatographic strip test for rapid diagnosis of malaria. Na-tional Institute of Parasitic Disease, Chinese Center for Disease Control and Prevention, WHO Collaborating Center For Malaria, Schis-tosomiasis and Filariasis. 2007; 25(5):415-418. 10. Liu Y, Du W, Zhen K, Fu L, Chen M. Com-parative study on rapid Dot-immunogold stain-ing and two immunogold silver staining assays for diagnosing schistosomiasis japonica. Southeast Asian J Trop Med Public Health. 2005;36(1):79-82.

Zhang GP, Guo JQ, Wang XN, Yang JX, Yang YY, Li QM, Li XW, Deng RG, Xiao ZJ, Yang JF, Xing GX, Zhao D. Development and evaluation of an immunochromatographic strip for trichinellosis detection. Vet Parasitol. 2006;137:286-293.

Oriol R, Williams JF, Perez-Esandi MV, Oriol C. Purification of lipoprotein antigens of Echi-nococcus granulosus from sheep hydatid cyst. Am J Trop Med Hyg. 1971;20:569-574.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Annual Biochem. 1976;72:248-254.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacterio-phage T4. Nature. 1970;227:680-685.

Benito A, Carmena D, Spinelli P, Postigo I, Martinez J, Estibalez JJ, Martin de la Cuesta F, Guisantes JA. The serological diagnosis of ca-nine echinococcosis by an enzyme immunoas-say useful for epidemiological surveys. Res Rev Parasitol. 2001;61:17-23.

Frens G. Controlled nucleation for the regula-tion of the particle size in monodisperse gold suspensions. Nature Phys Sci. 1976;241:20.

Kilic S, Babur C, Taylan OA. Comparison of the results of indirect hemagllutination and ELISA methods for the cases prediagnosed as hydatid cyst disease. Microbiol Bull. 2007;41(4):571-577.

Smits HL, Eapen CK, Sugathan S, Kuriakose M, Gasem MH, Yersin C, Sasaki D, Pujianto B, Vestering M, Abdoel TH, Gussenhoven GC. Lateril-flow assay for rapid serodiagnosis of human leptospirosis. Clin Diag Lab Immu-nol. 2001;8:166-169.

Liu YS, Du WP, Wu ZX. Dot-immunogold-silver staining in the diagnosis of cysticercosis. Int J Parasitol. 1996;26:127-129.

Chemale G, Ferreira HB, Barret J, Brophy PM, Zaha A. Echinococcus granulosus antigen B hydro-phobic ligand binding properties. Biochem Bi-ophys Acta Trop. 2005;47:189-194.

Tawfeek GM, Elwakil HS, Awad NS, El-Ho-seiny L, Thabet HS, Sarhan RM, Darweesh SK, Anwar WA. Genetic variability of antigen B among Echinococcus granulosus Egyptian isolates. Korean J Parasitol. 2009;47(3):259 -264.

Sadjjadi SM, Abidi H Sarkari B, Izadpanah A, Kazemian S. Evaluation of enzyme linked im-munosorbent assay utilizing native antigen B for serodiagnosis of human hydatidosis. Iranian J Parasitol. 2007;4(3):167-172.

Wu ZX, Du WP, Liu YS, Rong YW, Yuan SY, Ji XH. Comparative studies of Dot-IGSS and Dot-ELISA in detection antibodies from sera of schistosomiasis patients. Chinese Schisto-somiasis Cont. 1991;3:26-27. 24. Liu YS, Zeng KY, Chen M, Linlin F, Wenping D, Zhixu S. Study on detecting antibodies to Toxoplasma Gondii in pooled serum of blood donors by Dot-IGSS. Southeast Asian J Trop Med Publ Health. 2001;32(3):558 -561. 25. Wu ZX, Du WP, Rong YW, Ji XH, Yuan SY. The use of dot-immunogold-silver staining (Dot-IGSS), Dot-ELISA and Dot-IGSS to de-tect serum antibodies from clonorchiasis pa-tients: a comparative study. Southeast Asian J Trop Med Publ Health. 1993;24(4):677-679.

How to Cite
Jahani Z, Meshgi B, Rajabi-Bzl M, Jalousian F, Hasheminasab S. Improved serodiagnosis of hydatid cyst disease using gold nanoparticle labeled antigen B in naturally infected sheep. IJPA. 9(2):218-25.