Evaluation of Dot Immunogold Filtration Assay (DIGFA) By Recombinant Protein EPC1 for Anti- Echinococcus granulosus IgG Antibody.

  • Somayeh Kordafshari Dept. of Parasitology, School of Veterinary Medicine, Tehran University, Tehran, Iran.
  • Seyed Hossein Hosseini Dept. of Parasitology, School of Veterinary Medicine, Tehran University, Tehran, Iran.
  • Fatemeh Jalousian Dept. of Parasitology, School of Veterinary Medicine, Tehran University, Tehran, Iran.
  • Masoumeh Rajabibazl Dept. of Clinical Biochemistry, School of Medicine, Shahid Beheshti University, Tehran, Iran.
  • Malcolm Jones Dept. of Parasitology, School of Veterinary Sciences, Queensland University, Queensland, Australia.
  • Fazeleh Etebar Dept. of Parasitology, School of Veterinary Medicine, Tehran University, Tehran, Iran.
Keywords: Dot immunogold filtration assay, Echinococcus granulosus, Recombinant EPC1, Serodiagnosis


 Background: Cystic echinococcosis is an important zoonosis caused by the Echinococcus granulosus with a substantial impact in human and animal health in en-demic areas. The purpose of the present study was serodiagnosis optimizing of dog echinococcosis in order to achieve a rapid diagnostic method. Methods:Eight dogs were challenged with protoscoleces in order to have posi-tive echinococcosis serum and 2 two-month old puppies were used as uninfected controls. Colloidal gold was prepared by controlled reduction of a boiling solu-tion of chloroauric acid (H [AuCl4]) with sodium citrate and labeled with recom-binant EPC1. Dot immunogold filtration assay (DIGFA) was developed by coat-ing rEPC1 labeled colloidal gold on nitrocellulose membrane. The canine sera, taken three times including, 15, 28 and 35 days post infection were tested. A total of 30 serum samples including 24 sera from 8 infected dogs and 6 sera from 2 puppies were comparatively detected with both DIGFA and ELISA. Results:Gold labeled antigen, showed a dark purple dot with agglutination parti-cles in positive sera and light purple dot without agglutination in negative sera. Among 30 serum samples, 23 were positive and 7 were negative with DIGFA and 24 were positive, 6 were negative with ELISA. Conclusion:DIGFA as a rapid and simple procedure could be utilized in quickly diagnosis of echinococcosis.


Rajabloo M, Hosseini SH, Jalousian F. Morphological and molecular characterisation of Echinococcus granulosus from goat isolates in Iran. Acta Trop. 2012; 123:67-71.

Rokni MB. Echinococcosis /hydatidosis in Iran. Iran J Parasitol. 2009; 4(2):1-16.

Moro P, Schantz PM. Echinococcosis: a review. Int J Infect Dis. 2009; 13:125-33.

Brunetti E, Garcia HH, JunghanssT. International CE Workshop in Lima, Peru, Cystic echinococcosis: chronic, complex, and still neglected. PLoSNegl Trop Dis. 2011; 5:11-46.

Ahmadi NA, Badi F. Human hydatidosis in Tehran, Iran: A retrospective epidemiological study of surgical cases between 1999 and 2009 at two university medical centers. Trop Biomed. 2011; 28(2):450-6.

Tavakoli H, Bayat M, Kousha A. Hydatidosis Infection Study in Human and Livestock Populations During 2002-2007. Am-Eurasian J Agri Environ Sci. 2008; 4:473-7.

Amin Pour A, Hosseini SH, Shayan P. Comparative genotyping of Echinococcus granulosus infecting buffalo in Iran using cox1 gene. Parasitol Res. 2011; 108:1229-34.

WHO Informal Working Group. International classification of ultrasound images in cystic echinococcosis for application in clinical and field epidemiological settings. Acta Trop. 2003; 85:253-61.

Carmena D, Benito A, Eraso E. Antigens for the immunodiagnosis of Echinococcus granulosus infection. An update. Acta Trop. 2006; 98:74-86.

Chen XH, Wen H, Zhang ZX, Feng XH, Zhang JP, Zhang JH, Ma XD, Zheng SS. Field trial on rapid detection of echinococcosis by dot immunogold filtration assay (DIGFA) with whole blood sample. Chin J Parasitol Parasit Dis. 2005; 23(2):90-92.

Liu DY, Hu WQ, Zhang HM. Application of dot immunogold filtration assay for detecting serum antibodies in clonorchiasis patients. Chin J Parasitol Parasit Dis .2001a;19(2):97-99.

Liu ZP, Li H, Zhu LY. Application of dot immunogold filtration assay in antibody detection for cysticercosis. Chinese Journal of Parasitology & Parasitic Diseases. 2001b; 19(6):354-6.

Wang Y, Shim, XH, Gan XX. Dot immuno-gold filtration assay in the diagnosis of suspected paragonimiasis and evaluation of chemotherapeutic effect. Chin J Parasitol Parasit Dis. 2007; 25(1):65-68.

Abdi J, Kazemi B, Haniloo A, Mohebali M, Mahmoudi M, Rezaei S, Bandehpour M, Maghen L, Rokni MB. Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus granulosus for Diagnosis of Human Hydatidosis. Iran J Parasitol. 2010; 5(3):1-10.

Bora U, Chugh L, Nahar P. Covalent im- mobilization of proteins onto photoacti- vated polystyrene microtiter plates for enzyme linked immunosorbent assay procedures. J Immunol Meth. 2002; 268:171-7.

Frens G. Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions. Nat Phys Sci. 1973; 241:20-22.

Rance G, Marsh D, Khlobystov A. Extinction coefficient analysis of small alkanethiolate-stabilised gold nanoparticles. Chem Phys Lett. 2008; 460:230-6.

Etebar F, Jalousian F, Hosseini SH, Kordafshari S, Najafi A. Immunoproteomics approach for EPC1 antigenic epitope prediction of G1 and G6 strains of Echinococcus granulosus. Parasitol Res. 2013; 112(9):3129-35.

Dar VS, Ghosh S, Broor S. Rapid detection of rotavirus by using colloidal gold particles labeled with monoclonal antibody. J Virol Meth. 1994; 47:51-58.

Xiao LY, Yan XJ, Mi MR, Han FC, Hou Y. Preliminary study of a dot immunogold filtration assay for rapid detection of anti-HCV IgG. World J Gastroenterol. 1999; 5(4):349-50.

Urdal P, Borch SM, Landaas S, Krutnes MB, Gogstad GO, Hjortdahl P. Rapid immunometric measurement of C-Reactive protein in whole blood. Clin Chem. 1992; 38:580-4.

Beristain CN, Rojkin LF, Lorenze LE. Evaluation of dipstick method for detection of human immunodeficiency virus infection. J Clin Lab Anal, 1995; 9:347-50.

Spielberg F, Kabeya CM, Ryder RW, Kifuani NK, Harris J, Bender TR. Field testing and comparative evaluation of rapid visually read screening assays for antibody to human

immunodeficiency virus. The Lancet. 1989; 1:580-4

Wu W, Xu DZ, Yan Y-P, Zhang JX, Liu Y, Li RL. Evaluation of dot immunogold filtration assay foranti-HAV IgM antibody. World J Gastroenterol. 1999; 5(2):132-4

Huang Q, Lan X, Tong T, Wu X, Chen M, Feng X, Liu R, Tang Y, Zhu Z. Dot-Immunogold Filtration Assay as a Screening Test for Syphilis. J Clin Microbiol. 1996; 34(8): 2011-13.

Ye Y, Zhou Y, Mo Z, Cheng W, Yang S, Wang X, Chen F. Rapid detection of aflatoxin B(1) on membrane by dot-immunogold filtration assay. Talanta. 2010; 81(3):792-8.

Wang X, Zhan W, Xing J. Development of dot-immunogold filtration assay to detect white spot syndrome virus of shrimp.J Virol Meth. 2006; 132: 212-5.

How to Cite
Kordafshari S, Hosseini SH, Jalousian F, Rajabibazl M, Jones M, Etebar F. Evaluation of Dot Immunogold Filtration Assay (DIGFA) By Recombinant Protein EPC1 for Anti- Echinococcus granulosus IgG Antibody. IJPA. 10(1):30-.