Original Article

Evaluation of the Ability of Antigen B Originated from Echinococcus granulosus Sensu Stricto and E. canadensis for the Diagnosis of Confirmed Human Cystic Echinococcosis Using ELISA


Background: Cystic echinococcosis (CE) is an important zoonotic parasitic disease worldwide. Application of species-specific antigen for serodiagnosis of human CE has not been utilized, so far. In this regard, AgB originated from Echinococcus granulosus sensu stricto (G1-G3) and E. canadensis (G6/G7) CE cysts, confirmed by molecular biology and sequencing was used for evaluation of their ability in the diagnosis of confirmed human CE.

Methods: The hydatid cyst fluid (HCF) of E. granulosus sensu stricto and E. canadensis species were separately, used for preparation of AgB during 2017-2018 in Shiraz and Tehran, Iran. A total of 45 sera samples from confirmed CE patients, 102 sera from healthy people as negative control and 44 sera from other parasitic diseases, were used for measurement of the diagnostic ability of antigen B originated from E. granulosus sensu stricto and E. canadensis species of CE, alone or in 50%:50% mixture using ELISA method.

Results: Overall, 38 (84.4%) out of 45 confirmed human CE were positive by ELISA using AgB originated from E. granulosus sensu stricto. This items for AgB originated from E. canadensis was 39 (86.6) out of 45 serum samples. A total of 39 out of 45 samples (86.6%) showed positivity by a mixture of antigen B originating from both species. The specificity of the above tests was calculated as 93.15%, 96.58%, and 93.84%, respectively.

Conclusion:  Due to the diversity of the cyst species in human population, application of AgB from prevalent species alone or in combination with other species is suggested.

1. Thompson RCA. Biology and systematics of Echinococcus. Adv Parasitol. 2017; 95:65–109.
2. Vuitton DA, McManus DP, Rogan MT, et al. International consensus on terminology to be used in the field of echinococcosis. Parasite. 2020; 27:41.
3. Dalimi A, Mobedi I. Helminth parasites of carnivores in northern Iran. Ann Trop Med Parasitol. 1992; 86(4):395–7.
4. Zibaei M, Azargoon A, Ataie-Khorasgani M, et al. The serological study of cystic echinococcosis and assessment of surgical cases during 5 years (2007-2011) in Khorram Abad, Iran. Niger J Clin Pract. 2013; 16(2):221-5.
5. Eckert J, Gemmell MA, Meslin FX, et al. WHO/OIE manual on echinococcosis in humans and animals: a public health prob-lem of global concern, 2001. Paris.
6. Sadjjadi SM. Present situation of echino-coccosis in the Middle East and Arabic North Africa. Parasitol Int. 2006; 55 Suppl, S197-202.
7. Rokni MB. Echinococcosis/hydatidosis in Iran. Iran J Parasitol. 2009; 4(2):1-16.
8. Deplazes P, Rinaldi L, Alvarez Rojas CA, et al. Global distribution of alveolar and cystic echinococcosis. Adv Parasitol. 2017; 95:315–493.
9. Fasihi Harandi M, Budke CM, Rostami S. The monetary burden of cystic echinococ-cosis in Iran. PLoS Negl Trop Dis. 2012; 6(11):e1915.
10. Narra R, Maestri M, Budke CM, et al. Costs associated with surgically treated cases of abdominal cystic echinococcosis: a single center's experience from 2008 to 2014, Pavia, Italy. Am J Trop Med Hyg. 2016; 95(2):405-9.
11. Brunetti E, Kern P, Vuitton DA. Expert consensus for the diagnosis and treatment of cystic and alveolar echinococcosis in humans. Acta Trop. 2010; 114(1):1-16.
12. Sadjjadi SM, Ardehali S, Noman-Pour B, et al. Diagnosis of cystic echinococcosis: ul-trasound imaging or countercurrent im-munoelectrophoresis? East Mediterr Health J. 2001; 7(6):907-11.
13. Fratini F, Tamarozzi F, Macchia G, et al. Proteomic analysis of plasma exosomes from Cystic Echinococcosis patients pro-vides in vivo support for distinct immune response profiles in active vs inactive in-fection and suggests potential biomarkers. PLoS Negl Trop Dis. 2020; 5; 14(10):e0008586.
14. Khalilpour A, Sadjjadi SM, Moghadam ZK, et al. Lateral flow test using Echinococcus granulosus native antigen B and comparison of IgG and IgG4 dipsticks for detection of human cystic echinococcosis. Am J Trop Med Hyg. 2014; 91(5):994-9.
15. Wen H, Vuitton L, Tuxun T, et al. Echino-coccosis: advances in the 21st century. Clin Microbiol Rev. 2019; 32(2):e00075-18.
16. Sadjjadi FS, Ahmadi N, Rezaie-Tavirani M, et al. Following up of Surgical Treated Human Liver Cystic Echinococcosis: A Proteomics Approach. Iran J Parasitol. 2021; 16(1):11-22.
17. Carmena D, Benito A, Eraso E. Antigens for the immunodiagnosis of Echinococcus granulosus infection: an update. Acta Trop. 2006; 98(1):74-86.
18. Folle AM, Kitano ES, Lima A, et al. Char-acterisation of antigen B protein species present in the hydatid cyst fluid of Echino-coccus canadensis G7 genotype. PLoS Negl Trop Dis. 2017; 11(1):e0005250.
19. Muzulin PM, Kamenetzky L, Gutierrez AM, et al. Echinococcus granulosus antigen B gene family: further studies of strain poly-morphism at the genomic and transcrip-tional levels. Exp Parasitol. 2008; 118(2):156-64.
20. Mohammadzadeh T, Sako Y, Sadjjadi SM, et al. Comparison of the usefulness of hy-datid cyst fluid, native antigen B and re-combinant antigen B8/1 for serological diagnosis of cystic echinococcosis. Trans R Soc Trop Med Hyg. 2012; 106(6):371-5.
21. Romig T, Ebi D, Wassermann M. Taxon-omy and molecular epidemiology of Echi-nococcus granulosus sensu lato. Vet Parasitol. 2015; 213(3-4):76-84.
22. Sadjjadi SM, Mikaeili F, Karamian M, et al. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans. Int J Parasitol. 2013; 43(11):875-7.
23. Alvarez Rojas CA, Romig T, Lightowlers MW. Echinococcus granulosus sensu lato geno-types infecting humans--review of current knowledge. Int J Parasitol. 2014; 44(1):9-18.
24. Yanagida T, Mohammadzadeh T, Kam-hawi S, et al. Genetic polymorphisms of Echinococcus granulosus sensu stricto in the Middle East. Parasitol Int. 2012; 61(4):599-603.
25. Mardani P, Ezabadi AT, Sedaghat B, et al. Pulmonary hydatidosis genotypes isolates from human clinical surgery based on se-quencing of mitochondrial genes in Fars, Iran. J Cardiothorac Surg. 2021; 16(1):167.
26. Ebrahimipour M, Sadjjadi SM, Yousofi Darani H, et al. Molecular studies on cystic echinococcosis of camel (Camelus dromedari-us) and report of Echinococcus ortleppi in Iran. Iran J Parasitol. 2017; 12(3):323-331.
27. Bowles J, Blair D, McManus DP. Genetic variants within the genus Echinococcus identi-fied by mitochondrial DNA sequencing. Mol Biochem Parasitol. 1992; 54(2):165-73.
28. Bowles J, McManus DP. NADH dehy-drogenase 1 gene sequences compared for species and strains of the genus Echinococcus. Int J Parasitol. 1993; 23(7):969-72.
29. Hall T. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. In: Nucleic Acids Symp. 1999; 41,95-8.
30. Kearse M, Moir R, Wilson A, et al. Gene-ious Basic: an integrated and extendable desktop software platform for the organi-zation and analysis of sequence data. Bio-informatics. 2012; 28(12):1647-9.
31. Oriol R, Williams JF, Pérez Esandi MV, et al. Purification of lipoprotein antigens of Echinococcus granulosus from sheep hydatid fluid. Am J Trop Med Hyg. 1971; 20(4):569-74.
32. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54.
33. Sadjjadi SM, Abidi H, Sarkari B, et al. Eval-uation of enzyme linked immunosorbent assay, utilizing native antigen B for serodi-agnosis of human hydatidosis. Iran J Im-munol. 2007; 4(3):167-72.
34. MedCalc Software Ltd. Diagnostic test evaluation calculator. Available from: https://www.medcalc.org/calc/diagnostic_test.php (Version 20.011; accessed Sep-tember 18, 2021)
35. Pawłowski ZS, Eckert J, Vuitton DA, et al. Echinococcosis in humans: clinical aspects, diagnosis and treatment. WHO/OIE manual on echinococcosis in humans and animals: a public health problem of global concern. 2001: 20-66.
36. Sedaghat B, Hajjaran H, Sadjjadi FS, et al. Proteomic characterization of hydatid cyst fluid: two-dimensional electrophoresis (2-DE) setup through optimizing protein ex-traction. BMC Res Notes. 2021; 11; 14(1):22.
37. Lightowlers MW, Rickard MD, Honey RD, et al. Serological diagnosis of Echinococcus granulosus infection in sheep using cyst fluid antigen processed by antibody affinity chromatography. Aust Vet J. 1984; 61(4):101-8.
38. Gottstein B, Schantz PM, Todorov T, et al. An international study on the serological differential diagnosis of human cystic and alveolar echinococcosis. Bull World Health Organ. 1986; 64(1):101-5.
39. Sadjjadi FS, Rezaie-Tavirani M, Ahmadi NA, et al. Proteome evaluation of human cystic echinococcosis sera using two di-mensional gel electrophoresis. Gastroen-terol Hepatol Bed Bench. 2018; 11(1):75-82.
40. Rahimi H, Sadjjadi S, Sarkari B. Perfor-mance of antigen B isolated from different hosts and cyst locations in diagnosis of cystic echinococcosis. Iran J Parasitol. 2011; 6(1):12-9.
41. Verastegui M, Moro P, Guevara A, et al. Enzyme-linked immunoelectrotransfer blot test for diagnosis of human hydatid disease. J Clin Microbiol. 1992; 30(6):1557-61.
42. Wattal C, Malla N, Khan IA, et al. Com-parative evaluation of enzyme-linked im-munosorbent assay for the diagnosis of pulmonary echinococcosis. J Clin Micro-biol. 1986; 24(1):41-6.
43. Force L, Torres JM, Carrillo A, et al. Eval-uation of eight serological tests in the di-agnosis of human echinococcosis and fol-low-up. Clin Infect Dis. 1992; 15(3):473-80.
44. Babba H, Messedi A, Masmoudi S, et al. Diagnosis of human hydatidosis: compar-ison between imagery and six serologic techniques. Am J Trop Med Hyg. 1994; 50(1):64-8.
45. Gottstein B, Eckert J, Michael SA, et al. Echinococcus granulosus antigens: immunoelec-trophoretic and western blot analysis of hydatid cyst fluids. Parasitol Res. 1987; 73(2):186-9.
46. Williams JF, Pérez Esandi MV, Oriol R. Evaluation of purified lipoprotein antigens of Echinococcus granulosus in the immunodi-agnosis of human infection. Am J Trop Med Hyg. 1971; 20(4):575-9.
47. Sadjjadi SM, Ebrahimipour M, Sadjjadi FS. Comparison between Echinococcus granulosus sensu stricto (G1) and E. canadensis (G6) mitochondrial genes (cox1 and nad1) and their related protein models using experi-mental and bioinformatics analysis. Com-put Biol Chem. 2019; 79:103-109.
IssueVol 17 No 3 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v17i3.10626
Cystic echinococcosis Hydatidosis Species-specific antigens Diagnosis Human

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How to Cite
Sadjjadi SF, Mohammadzadeh T, Hafezi F, Sadjjadi SM. Evaluation of the Ability of Antigen B Originated from Echinococcus granulosus Sensu Stricto and E. canadensis for the Diagnosis of Confirmed Human Cystic Echinococcosis Using ELISA. Iran J Parasitol. 2022;17(3):358-365.