Hostname: page-component-cd9895bd7-lnqnp Total loading time: 0 Render date: 2024-12-18T14:16:09.387Z Has data issue: false hasContentIssue false

Comparison of parasite loads in serum and blood samples from patients in acute and chronic phases of Chagas disease

Published online by Cambridge University Press:  17 April 2018

Carolina Hernández
Affiliation:
Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad el Rosario, Bogotá, Colombia
Aníbal Teherán
Affiliation:
Grupo de investigación COMPLEXUS, Fundación Univeresitarias Juan N. Corpas, Bogotá, Colombia Medicina de Emergencias, Escuela de Medicina y Ciencias de la Salud, Universidad del Rosario, Bogotá, Colombia
Carolina Flórez
Affiliation:
Grupo de Parasitología, Instituto Nacional de Salud, Bogotá, Colombia
Juan David Ramírez*
Affiliation:
Grupo de Investigaciones Microbiológicas-UR (GIMUR), Programa de Biología, Facultad de Ciencias Naturales y Matemáticas, Universidad el Rosario, Bogotá, Colombia
*
Author for correspondence: Juan David Ramírez, E-mail: [email protected]

Abstract

Molecular methods have been developed for the detection and quantification of Trypanosoma cruzi DNA in blood samples from patients with Chagas disease. However, aspects of sample processing necessary for quantitative real-time PCR (qPCR), such as the addition of guanidine hydrochloride to whole blood samples, may limit timely access to molecular diagnosis. We analysed 169 samples from serum and guanidine-EDTA blood (GEB) obtained from patients in acute and chronic phases of Chagas disease. We applied qPCR targeted to the satellite DNA region. Finally, we compared the parasite loads and cycle of threshold values of the qPCR. The results confirmed the usefulness of serum samples for the detection and quantification of parasite DNA in patients with Chagas disease, especially in the acute phase. However, the parasite loads detected in serum samples from patients in the chronic phase were lower than those detected in GEB samples. The epidemiological implications of the findings are herein discussed.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bern, C, Longo, DL and Bern, C (2015) Chagas’ disease. New England Journal of Medicine 373, 456466.Google Scholar
Bianchi, F, et al. (2015) Follow-up of an asymptomatic Chagas disease population of children after treatment with nifurtimox (Lampit) in a sylvatic endemic transmission area of Colombia. PLoS Neglected Tropical Diseases 9, e0003465.Google Scholar
Brasil, PEAA, et al. (2010) ELISA versus PCR for diagnosis of chronic Chagas disease: systematic review and meta-analysis. BMC Infectious Diseases 10, 337.Google Scholar
Duffy, T, et al. (2009) Accurate real-time PCR strategy for monitoring bloodstream parasitic loads in Chagas disease patients. PLoS Neglected Tropical Diseases 3, e419.Google Scholar
Duffy, T, et al. (2013) Analytical performance of a multiplex real-time PCR assay using TaqMan probes for quantification of Trypanosoma cruzi satellite DNA in blood samples. PLoS Neglected Tropical Diseases 7, e2000.Google Scholar
Cucunubá, ZM, et al. (2016) Increased mortality attributed to Chagas disease: a systematic review and meta-analysis. Parasites and Vectors 9, 42.Google Scholar
Feilij, H, Muller, L and Gonzalez Cappa, SM (1983) Direct micromethod for diagnosis of acute and congenital Chagas’ disease. Journal of Clinical Microbiology 18, 327330.Google Scholar
Filigheddu, MT, Górgolas, M and Ramos, JM (2017) Orally-transmitted Chagas disease. Medica Clínica 148, 125131.Google Scholar
Hernandez, C and Ramirez, JD (2013) Molecular diagnosis of vector-borne parasitic diseases. Air Water Borne Diseases 2, 110.Google Scholar
Hernández, C, et al. (2016 a) Molecular diagnosis of Chagas disease in Colombia: parasitic loads and discrete typing units in patients from acute and chronic phases. PLoS Neglected Tropical Diseases 10, e0004997.Google Scholar
Hernández, C, et al. (2016 b) High-resolution molecular typing of Trypanosoma cruzi in 2 large outbreaks of acute Chagas disease in Colombia. Journal of Infectious Diseases 214, 12521255.Google Scholar
López, MC, et al. (1999) Inmunodiagnóstico de la infección chagásica por ELISA. Biomédica 19, 159163.Google Scholar
Melo, MF, et al. (2015) Usefulness of real time PCR to quantify parasite load in serum samples from chronic Chagas disease patients. Parasites and Vectors 8, 154.Google Scholar
Moreira, OC, et al. (2013) Towards the establishment of a consensus real-time qPCR to monitor Trypanosoma cruzi parasitemia in patients with chronic Chagas disease cardiomyopathy: a substudy from the BENEFIT trial. Acta Tropica 125, 2331.Google Scholar
Morillo, CA, et al. (2015) Randomized trial of benznidazole for chronic Chagas’ cardiomyopathy. New England Journal of Medicine 373, 112.Google Scholar
Ramírez, JC, et al. (2015) Analytical validation of quantitative real-time PCR methods for quantification of Trypanosoma cruzi DNA in blood samples from Chagas disease patients. Journal of Molecular Diagnostics 17, 605615.Google Scholar
Ramírez, JD, et al. (2013) Molecular epidemiology of human oral Chagas disease outbreaks in Colombia. PLoS Neglected Tropical Diseases. 7, 17.Google Scholar
Russomando, G, et al. (1992) Polymerase chain reaction-based detection of Trypanosoma cruzi DNA in serum. Journal of Clinical Microbiology 30, 28642868.Google Scholar
Russomando, G, et al. (1998) Treatment of congenital Chagas’ disease diagnosed and followed up by the polymerase chain reaction. American Journal of Tropical Medicine and Hygiene 59, 487491.Google Scholar
Shikanai-Yasuda, MA and Carvalho, NB (2012) Oral transmission of Chagas disease. Clinical Infectious Diseases 54, 845852.Google Scholar
Supplementary material: File

Hernández et al. supplementary material

Appendix 1

Download Hernández et al. supplementary material(File)
File 23.4 KB
Supplementary material: File

Hernández et al. supplementary material

Table S1

Download Hernández et al. supplementary material(File)
File 13.3 KB