AN EVALUATION STUDY OF ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) USING RECOMBINANT PROTEIN GRA1 FOR DETECTION OF IgG ANTIBODIES AGAINTS TOXOPLASMA GONDII INFECTIONS

Nina Difla Muflikhah, Wayan Tunas Artama

= http://dx.doi.org/10.20473/ijtid.v6i5.5903
Abstract views = 423 times | views = 1483 times

Abstract


Toxoplasmosis is an infectious disease caused by Toxoplasma gondii, an intracellular protozoan parasite that live inside the cells of the reticulo endothelial and parenchymal cells of human and animals (mammals and birds). Some cases of toxoplasmosis usually have no symptoms, but in any cases caused severe symptoms, such as hydrocephalus, microcephalus, intracranial calcification, retinal damage, brain abscess, mental retardation, lymphadenopathy, and others. Its severe symptoms usually showed a long time after first exposure, except symptoms showed by congenital transmission caused by infected mother. Early diagnosis is important to prevent the illness but methods for toxoplasmosis screening are still too expensive for developing country. Enzyme-linked immunosorbent assay (ELISA) allow the testing of a large number samples within short time frame and based on antibody or antigen detection. This study aimed to know the sensitivity and specificity of recombinat protein GRA1 as antigen using ELISA methods. We tested the sensitivity and spesificity of GRA1 protein as antigen in ELISA methods to diagnose toxoplasmosis and compared with ELISA Kit Commercial. Reliable laboratory testing is important to detect Toxoplasma gondii infection, and focused to improving the low cost and easy-to-use diagnostic instrument. Seventy sera collected and tested using both indirect ELISA, commercial ELISA kit and GRA1 protein coated as antigen. Fourty eight and fifty one samples showed positive IgG antibody result of ELISA-GRA1 and ELISA kit. Negative sample tested by ELISA-GRA1 was 22 samples and 19 sample tested by ELISA Kit. The sensitivity and specificity of GRA1-based on ELISA were 100% and 86.36%, positive prediction value (ppv) was 94.11%. These data indicate that the recombinant protein GRA1 is a highly immunogenic protein in human toxoplasmosis and become a promising marker for the screening of toxoplasmosis.


Keywords


Toxoplasmosis; recombinant protein GRA1; ELISA; Sensitivity; Specificity

Full Text:

PDF

References


Montoya JG, Liesenfeld O. Toxoplasmosis. Lancet (London, England). 2004 Jun 12;363(9425):1965–76.

Sibley LD. Toxoplasma gondii: perfecting an intracellular life style. Traffic. 2003 Sep;4(9):581–6.

Aubert D, Maine GT, Villena I, Hunt JC, Howard L, Sheu M, et al. Recombinant antigens to detect Toxoplasma gondii-specific immunoglobulin G and immunoglobulin M in human sera by enzyme immunoassay. J Clin Microbiol. 2000 Mar;38(3):1144–50.

Hassi A, Muller WA, Aspock H. An identical epitope inPneumocystis carinii andToxoplasma gondii causing serological cross reactions. Parasitol Res. 1991;77(4):351–2.

Cesbron-Delauw M-F, Lecordier L, Mercier C. Role of Secretory Dense Granule Organelles in the Pathogenesis of Toxoplasmosis. In: Toxoplasma gondii. Berlin, Heidelberg: Springer Berlin Heidelberg; 1996. p. 59–65.

Vercammen M, Scorza T, Huygen K, De Braekeleer J, Diet R, Jacobs D, et al. DNA vaccination with genes encoding Toxoplasma gondii antigens GRA1, GRA7, and ROP2 induces partially protective

immunity against lethal challenge in mice. Infect Immun. 2000 Jan;68(1):38–45.

Scorza T, D’Souza S, Laloup M, Dewit J, De Braekeleer J, Verschueren H, et al. A GRA1 DNA vaccine primes cytolytic CD8(+) T cells to control acute Toxoplasma gondii infection. Infect Immun. 2003 Jan;71(1):309–16.

Sulistyaningsih E, Moeljopawiro S, Subandono J, Artama WT. Cloning of cDNA Encoding GRA1 Protein of Tachyzoite Toxoplasma Gondii Local Isolate. Indones J Biotechnol. 2005;10(1):763–7.

Sibley LD, Mordue DG, Su C, Robben PM, Howe DK. Genetic approaches to studying virulence and pathogenesis in Toxoplasma gondii. Philos Trans R Soc Lond B Biol Sci. 2002 Jan 29;357(1417):81–8.

Carruthers VB. Host cell invasion by the opportunistic pathogen Toxoplasma gondii. Acta Trop. 2002 Feb;81(2):111–22.

Hiszczynska-Sawicka E, Kur J, Pietkiewicz H, Holec L, Gsior A, Myjak P. Efficient production of the Toxoplasma gondii GRA6, p35 and SAG2 recombinant antigens and their applications in the serodiagnosis of toxoplasmosis. Acta Parasitológica. 2005;50(3):249–54.

Retmanasari A, Widartono BS, Wijayanti MA, Artama WT. Analisis spasial dan faktor risiko toksoplasmosis di jawa tengah bagian selatan. Universitas Gadjah Mada; 2015.

Utami WS. Isolasi, karakterisasi dan uji diagnostik protein rekombinan GRA-1 takizoit toxoplasma gondii isolat lokal. Universitas Gadjah Mada; 2009.

Widayanti E. Subkloning dan over ekspresi gen penyandi protein GRA-1 Takizoit toxoplasma gondii isolat lokal. Universitas Gadjah Mada; 2008.

Wu X-N, Lin J, Lin X, Chen J, Chen Z-L, Lin J-Y. Multicomponent DNA vaccine-encoding Toxoplasma gondii GRA1 and SAG1 primes: anti-Toxoplasma immune response in mice. Parasitol Res. 2012 Nov 27;111(5):2001–9.

Black MW, Boothroyd JC. Lytic cycle of Toxoplasma gondii. Microbiol Mol Biol Rev. 2000 Sep;64(3):607–23.

Miller CM, Boulter NR, Ikin RJ, Smith NC. The immunobiology of the innate response to Toxoplasma gondii. Int J Parasitol. 2009 Jan;39(1):23–39.

Nam H-W. GRA proteins of Toxoplasma gondii: maintenance of hostparasite interactions across the parasitophorous vacuolar membrane. Korean J Parasitol. 2009 Oct;47 Suppl:S29-37.

Tizard I. Veterinary Immunology. 9th ed. Philadelphia (Pennsylvania): Saunders; 2012.

Döşkaya M, Kalantari-Dehaghi M, Walsh CM, Hiszczyńska-Sawicka E, Davies DH, Felgner PL, et al. GRA1 protein vaccine confers better immune response compared to codon-optimized GRA1 DNA vaccine. Vaccine. 2007 Feb 26;25(10):1824–37.

Holec-Gasior L. Toxoplasma gondii recombinant antigens as tools for serodiagnosis of human toxoplasmosis: current status of studies. Clin Vaccine Immunol. 2013 Sep;20(9):1343–51.

Subekti DT. Study of Antigenicity and Immunogenicity Gra1 Protein from Toxoplasma gondii. Indones Bull Anim Vet Sci. 2014 Apr 30;23(3).

Wang Z, Ge W, Huang S-Y, Li J, Zhu X-Q, Liu Q. Evaluation of recombinant granule antigens GRA1 and GRA7 for serodiagnosis of Toxoplasma gondii infection in dogs. BMC Vet Res. 2014 Apr 30;10(1):158.

Zhou H, Min J, Zhao Q, Gu Q, Cong H, Li Y, et al. Protective immune response against Toxoplasma gondii elicited by a recombinant DNA vaccine with a novel genetic adjuvant. Vaccine. 2012 Feb 27;30(10):1800–6.

Ferrandiz J, Mercier C, Wallon M, Picot S, Cesbron-Delauw M-F, Peyron F. Limited Value of Assays Using Detection of Immunoglobulin G Antibodies to the Two Recombinant Dense Granule Antigens, GRA1 and GRA6 Nt of Toxoplasma gondii, for Distinguishing between Acute and Chronic Infections in Pregnant Women. Clin Vaccine Immunol. 2004 Nov 1;11(6):1016–21.

Gedik Y, Gülçe İz S, Can H, Değirmenci Döşkaya A, İsmet Deliloğlu Gürhan S, Gürüz Y, et al. Immunogenic multistage recombinant protein vaccine confers partial protection against experimental toxoplasmosis mimicking natural infection in murine model. Trials Vaccinol. 2016;5:15–23.


Refbacks

  • There are currently no refbacks.




Copyright (c) 2017 Indonesian Journal of Tropical and Infectious Disease

View My Stats

IJTID Indexed by : 

                                                    

 


IJTID (pISSN 2085-1103eISSN 2356-0991is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Institute of Tropical Disease - Airlangga University
Gedung Lembaga Penyakit Tropis Lt.1, Kampus C Universitas Airlangga
Jln. Mulyorejo Surabaya 60115, Indonesia
E-mail: ijtid@itd.unair.ac.id