Serological Evaluation of EgAgB16 kDa, a Recombinant Antigen from Echinococcus granulosus for Diagnosis of Human Hydatidosis
-
J Abdi
,
B Kazemi
,
A Haniloo
,
M Mohebali
,
M Mahmoudi
,
S Rezaei
,
M Bandehpour
,
L Maghen
,
MB Rokni
Abstract
Background: Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test.
Methods: DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen.
Results: Recombinant protein was purified by affinity chromatography using His-Tag column. After purification, recombinant protein was confirmed by western blot analysis using His Tag monoclonal antibody or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16.
Conclusion: While the produced recombinant AgB16 kDa showed promising results in diagnosing human hydatidosis, but more investigations should be implemented to reach an accurate gold standard.
Torgerson PR, Deplazes P. Echinococcosis: diagnosis and diagnostic interpretation in population studies. Trends Parasitol. 2009; 25:164-70.
Rokni MB. Echinococcosis /hydatidosis in Iran. Iranian J Parasitol. 2009; 4:1-16.
Budke CM, Deplazes P, Torgerson PR. Global Socioeconomic Impact of Cystic Echinococcosis. Emerging Infectious Diseases. 2006; 12:296-303.
Sadjjadi SM. Present situation of echinococcosis in the Middle East and Arabic North Africa. Parasitol Int. 2006; 55 Suppl:S197-202.
Robert-Gangneux F, Tourte-Schaefer C. [Comparative value of 2 western blot techniques for confirmation of hydatidosis diagnosis]. Bull Soc Pathol Exot. 1999; 92:13-7. [In French].
Rogan MT, Craig PS, Zeyhle E, Romig T, Lubano GM, Deshan L. Evaluation of a rapid dot-ELISA as a field test for the diagnosis of cystic hydatid disease. Trans R Soc Trop Med Hyg. 1991; 85:773-7.
Rokni MB, Massoud J, Hanilo A. Comparison of adult somatic and cysteine proteinase antigens of Fasciola gigantica in enzyme linked immunosorbent assay for serodiagnosis of human fasciolosis. Acta Trop. 2003; 88:69-75.
Ramzy RM, Helmy H, El Zayyat EA, Rifaat MM, Abdel Hameed DM, Abdel- Baki MH. An enzyme-linked immunosorbent assay for detection of IgG1 antibodies specific to human cystic echinococcosis in Egypt. Trop Med Int Health. 1999; 4:616-20.
Sadjjadi SM, Ardehali S, Noman-Pour B, Kumar V, Izadpanah A. Diagnosis of cystic echinococcosis: ultrasound imaging or countercurrent immunoelectrophoresis? Eastern Mediterranean Health Journal. 2001; 7:907-11.
Kalantari E, Bandehpour M, Pazoki R, Taghipoor-Lailabadi N, Khazan H, Mosaffa N, et al. Application of recombinant Echinococcus granulosus antigen B to ELISA kits for diagnosing hydatidosis. Parasitology Research. 2010; 106:847-51.
Frosch P, Hartmann M, Mühschlegel F, Frosch M. Sequence heterogeneity of the echinococcal antigen B. Mol Biochem Parasitol. 1994; 64:171-5.
Rott MB, Fernandes V, Farias S, Ceni J, Ferreira HB, Haag KL, et al. Comparative analysis of two different sub units of AgB from Echinococcus granulosus; Gene sequence expression in E. coli and serological evaluation. Acta Trop. 2000; 75:331-4.
Shephered JC, Aitken A, McManus DP. A proatein secreted in vivo by Echinococcus granulosus, inhibits elastase, activity and neutrophil chemotaxis. Mol Biochem Parasitol. 1991; 44:81-90.
Rostami Nejad M, Nazemalhosseini Mojarad E, Nochi Z, Fasihi Harandi M, Cheraghipour K, Mowlavi GR, et al. Echinococcus granulosus strain differentiation in Iran based on sequence heterogeneity in the mitochondrial 12S rRNA gene. J Helminthol. 2008; 82:343-7.
Bandehpour M, Seyed N, Shadnoosh M, Pakzad P, Kazemi B. Using recombinant Chlamydia Major Outer Membrane Protein (MOMP) in ELISA diagnostic kit. Iran J Biotech. 2006; 4:239- 44.
Bora U, Chugh L, Nahar P. Covalent immobilization of proteins onto photoactivated polystyrene microtiter plates for enzyme- linked immunosorbent assay procedures. J Immunol Meth. 2002; 268:171-7.
Galen RS. Predictive values & efficiency of laboratory testing. Pediater Clin North Am. 1980; 27:861-689.
Akisu C, Delibas SB, Bicmen C, Ozkoc S, Aksoy U, Turgay N. Comparative evaluation of western blotting in hepatic and pulmonary cystic echinococcosis. Parasite. 2006; 13:321-6.
Baveja UK, Basak S, Thusoo TK. Immunodiagnosis of human hydatid disease. J Commun Dis. 1997; 29:313-9.
Ben Nouir N, Nun˜ez S, Gianinazzi C, Gorcii M, Mu¨ller N, Nouri A, et al. Assessment of Echinococcus granulosus somatic protoscolex antigens for serological follow-up of young patients surgically treated for cystic echinococcosis. J Clinic Microbiol 2008; 46:1631-40.
Biava MF, Dao A, Fortier B. Laboratory diagnosis of cystic hydatid disease. World J Surg. 2001; 25:10-4.
Rokni MB, Aminian B. Evaluation of the Enzyme-linked Immuno-electro Transfer Blot (EITB) technique using hydatid cyst antigens B/5 and total IgG antibodies in laboratory diagnosis of human hydatidosis. Pak J Med Sci. 2006; 22:127-31.
Sarkari B, Sadjjadi SM, Abidi H, Izadpana AH, Kazemian S, Rafati A. Application of= western blotting using native antigen B for serodiagnosis of human cystic echinococcosis. Iranian J Parasitol. 2007; 2:7-12.
Shi Z, Wang Y, Li Z, Li Z, Bo Y, Ma R, et al. Cloning, expression, and protective immunity in mice of a gene encoding the diagnostic antigen P-29 of Echinococcus granulosus. Acta Biochim Biophys Sin (Shanghai). 2009; 41:79-85.
Ben Nouir N, Gianinazzi C, Gorcii M, Muller N, Nouri A, Babba H, et al. Isolation and molecular characterization of recombinant Echinococcus granulosus P29 protein (recP29) and its assessment for the post-surgical serological follow-up of human cystic echinococcosis in young patients. Trans R Soc Trop Med Hyg. 2009; 103:355-64.
Hernandez-Gonzalez A, Muro A, Barrera I, Ramos G, Orduna A, Siles-Lucas M. Usefulness of four different Echinococcus granulosus recombinant antigens for serodiagnosis of unilocular hydatid disease (UHD) and postsurgical follow-up of patients treated for UHD. Clin Vaccine Immunol. 2008; 15:147-53.
Mamuti W, Yamasaki H, Sako Y, Nakao M, Xiao N, Nakaya K, et al. Molecular cloning, expression, and serological evaluation of an 8-kilodalton subunit of antigen B from Echinococcus multilocularis. J Clin Microbiol. 2004; 42:1082-8.
Rahimi HR, Kia EB, Mirhendi SH, Talebi A, Fasihi Harandi M, Jalalizand N, Rokni MB. A New Primer Pair in ITS1 Region for Molecular Studies on Echinococcus granulosus. Iranian J Publ Health. 2007;36 (1):45-49.
Abdi J, Kazemi B, Mohebali M, Bandehpour M, Rahimi M, Rokni MB. Gene cloning, expression, and serological evaluation of EgAgB12 kDa from Echinococcus granulosus. Ann Trop Med Parasitol. 2010.(Accepted, In press).
Virginio VG, Hernández A, Rott MB, Monteiro KM, Zandonai AF, Nieto A, et al. A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease. Clin Exp Immunol. 2003; 132:309-15.
de la Rue ML, Yamano K, Almeida CE, Iesbich MP, Fernandes CD, Goto A, et al. Serological reactivity of patients with Echinococcus infections (E. granulosus, E. vogeli, and E. multilocularis) against three antigen B subunits. Parasitol Res. 2010; 106:741-5.
Haniloo A, Massoud J, Rokni MB. Evaluation and comparison of antigen BELISA and antigen B-immunoblotting in immunodiagnosis of cystic hydatid disease. Pak J Med Sci. 2005; 21:352-6.
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| Issue | Vol 5 No 3 (2010) | |
| Section | Articles | |
| Keywords | ||
| Hydatidosis Echinococcus granulosus ELISA Recombinant Antigen | ||
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