Comparison of seven diagnostic tests to detect Trypanosoma cruzi infection in patients in chronic phase of Chagas disease
Objective: To compare the diagnostic performance of seven methods to determine Trypanosoma cruzi infection in patients with chronic Chagas disease.
Methods: Analytical study, using the case-control design, which included 205 people (patients with Chagasic cardiomyopathy, n= 100; control group, n= 105). Three enzyme linked immunosorbent assays, one indirect hemagglutination assay and one immunochromatographic test were assessed. Additionally, DNA amplification was performed via the PCR method using kinetoplast and nuclear DNA as target sequences. For the comparative analysis of diagnostic tests, the parameters used were sensitivity, specificity, positive and negative predictive values, Receiver Operator Characteristic (ROC), positive and negative likelihood ratio, as well as κ quality analysis.
Results: The commercial Bioelisa Chagas test showed the highest sensitivity (98%), specificity (100%), and positive and negative predictive values; additionally it had the highest discriminatory power. Otherwise, the amplification of T. cruzi DNA in blood samples showed low values of sensitivity (kinetoplast DNA= 51%, nuclear DNA= 22%), but high values of specificity (100%), and moderate to low discriminatory ability.
Conclusion: The comparative analysis among the different methods suggests that the diagnostic strategy of T. cruzi infection in patients with chronic Chagas disease can be performed using ELISA assays based on recombinant proteins and/or synthetic peptides, which show higher diagnosis performance and can confirm and exclude the diagnosis of T. cruzi infection. The molecular methods show poor performance when used in the diagnosis of patients with chronic Chagas disease.
- Chagas disease
- Trypanosoma cruzi
- Enzyme linked immunosorbent assay
- Recombinant Proteins
- Polymerase Chain Reaction
WHO . First WHO report on neglected tropical diseases. WHO; Geneva, Switzerland: 2010.
Rassi A, Jr, Rassi A, Marcondes de Rezende J American trypanosomiasis (Chagas disease) Infect Dis Clin North Am. 2012; 26(2): 275–91.
Moncayo A, Silveira AC. Current epidemiological trends for Chagas disease in Latin America and future challenges in epidemiology, surveillance and health policy. Mem Inst Oswaldo Cruz. 2009; 104(1): 17–30.
Gutierrez R, Angulo VM, Tarazona Z, Britto C, Fernandes O. Comparison of four serological tests for the diagnosis of Chagas disease in a Colombian endemic area. Parasitology. 2004; 129: 439–44.
Gomes YM, Lorena VM, Luquetti AO. Diagnosis of Chagas disease: what has been achieved? What remains to be done with regard to diagnosis and follow up studies? Mem Inst Oswaldo Cruz. 2009; 104(1): 115–21.
Pan American Health Organization Quantitative estimation of Chagas disease in the Americas. (OPS/HDM/CD/425-06). 2006.
Meira WS, Galvao LM, Gontijo ED, Machado-Coelho GL, Norris KA, Chiari E. Trypanosoma cruzi recombinant complement regulatory protein: a novel antigen for use in an enzyme-linked immunosorbent assay for diagnosis of Chagas’ disease. J Clin Microbiol. 2002; 40: 3735–40.
Pereira-Chioccola VL, Fragata-Filho AA, Levy AM, Rodrigues MM, Schenkman S. Enzyme-linked immunoassay using recombinant trans-sialidase of Trypanosoma cruzi can be employed for monitoring of patients with Chagas’ disease after drug treatment. Clin Diagn Lab Immunol. 2003; 10: 826–30.
da Silveira JF, Umezawa ES, Luquetti AO. Chagas disease: recombinant Trypanosoma cruzi antigens for serological diagnosis. Trends Parasitol. 2001; 17: 286–91.
Marcon GE, Andrade PD, de Albuquerque DM, Wanderley Jda S, de Almeida EA, Guariento ME, et al. Use of a nested polymerase chain reaction (N-PCR) to detect Trypanosoma cruzi in blood samples from chronic chagasic patients and patients with doubtful serologies. Diagn Microbiol Infect Dis. 2002; 43: 39–43.
Piron M, Fisa R, Casamitjana N, Lopez-Chejade P, Puig L, Verges M, et al. Development of a real-time PCR assay for Trypanosoma cruzi detection in blood samples. Acta Trop. 2007; 103: 195–200.
Deborggraeve S, Coronado X, Solari A, Zulantay I, Apt W, Mertens P, et al. T. cruzi OligoC-TesTa simplified and standardized polymerase chain reaction format for diagnosis of Chagas disease. PLoS Negl Trop Dis. 2009; 3(6): e450.
Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988; 16: 1215.
Orozco LC. Medición en Salud, Diagnóstico, Evaluación de Resultados.Un manual crítico más allá de lo básico. Bucaramanga, Colombia: Publicaciones UIS; 2010.
Kraemer H. Evaluating medical test. Objective and Quantitative Guidelines. Newbury Park, USA: Sage publications, Inc; 1992.
Umezawa ES, Bastos SF, Camargo ME, Yamauchi LM, Santos MR, Gonzalez A, et al. Evaluation of recombinant antigens for serodiagnosis of Chagas’ disease in South and Central America. J Clin Microbiol. 1999; 37: 1554–60.
Ramirez JD, Guhl F, Umezawa ES, Morillo CA, Rosas F, Marin-Neto JA, et al. Evaluation of adult chronic Chagas’ heart disease diagnosis by molecular and serological methods. J Clin Microbiol. 2009; 47: 3945–51.
Houghton RL, Benson DR, Reynolds LD, McNeill PD, Sleath PR, Lodes MJ, et al. A multi-epitope synthetic peptide and recombinant protein for the detection of antibodies to Trypanosoma cruzi in radioimmunoprecipitation-confirmed and consensus-positive sera. J Infect Dis. 1999; 179: 1226–34.
Cotrim PC, Paranhos-Baccala G, Santos MR, Mortensen C, Cano MI, Jolivet M, et al. Organization and expression of the gene encoding an immunodominant repetitive antigen associated to the cytoskeleton of Trypanosoma cruzi. Mol Biochem Parasitol. 1995; 71: 89–98.
Avila HA, Pereira JB, Thiemann O, De Paiva E, DeGrave W, Morel CM, et al. Detection of Trypanosoma cruzi in blood specimens of chronic chagasic patients by polymerase chain reaction amplification of kinetoplast minicircle DNA: comparison with serology and xenodiagnosis. J Clin Microbiol. 1993; 31: 2421–6.
Schijman AG, Bisio M, Orellana L, Sued M, Duffy T, Mejia Jaramillo AM, et al. International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients. PLoS Negl Trop Dis. 2011; 5(1): e931.
Avila HA, Sigman DS, Cohen LM, Millikan RC, Simpson L. Polymerase chain reaction amplification of Trypanosoma cruzi kinetoplast minicircle DNA isolated from whole blood lysates: diagnosis of chronic Chagas’ disease. Mol Biochem Parasitol. 1991; 48: 211–21.
Gonzalez A, Prediger E, Huecas ME, Nogueira N, Lizardi PM. Minichromosomal repetitive DNA in Trypanosoma cruzi: its use in a high-sensitivity parasite detection assay. Proc Natl Acad Sci USA. 1984;81: 3356–60.
Elias MC, Vargas NS, Zingales B, Schenkman S. Organization of satellite DNA in the genome of Trypanosoma cruzi. Mol Biochem Parasitol. 2003; 129: 1–9.
Gonzalez CI, Ortiz S, Solari A. Colombian Trypanosoma cruzi major genotypes circulating in patients: minicircle homologies by cross-hybridization analysis. Int J Parasitol. 2010; 40(14): 1685–92.
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