Monitoring the parasite load in chronic Chagas disease patients: comparison between blood culture and quantitative real time PCR

被引:22
|
作者
D'Avila, Daniella Alchaar [1 ]
Galvao, Lucia Maria C. [1 ]
Sousa, Giovane R. [2 ]
Britto, Constanca [3 ]
Moreira, Otacilio C. [3 ]
Chiari, Egler [1 ]
机构
[1] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Parasitol, Programa Posgrad Parasitol, Belo Horizonte, MG, Brazil
[2] Harvard Med Sch, Joslin Diabet Ctr, Sect Immunobiol, Boston, MA USA
[3] Fiocruz MS, Lab Biol Mol & Doencas Endem, Inst Oswaldo Cruz, Rio De Janeiro, RJ, Brazil
来源
PLOS ONE | 2018年 / 13卷 / 11期
关键词
POLYMERASE-CHAIN-REACTION; TRYPANOSOMA-CRUZI INFECTION; LINKED-IMMUNOSORBENT-ASSAY; CHRONIC PHASE; DIAGNOSIS; DNA; SAMPLES; QUANTIFICATION; IDENTIFICATION; AMPLIFICATION;
D O I
10.1371/journal.pone.0208133
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background Despite the improvements in diagnostic tools for detection of Trypanosoma cruzi in human blood samples, the isolation of parasite from bloodstream in the chronic phase of Chagas disease is challenging. Thus, there is an increasing interest in the development of strategies that allow an accurate monitoring of the parasite load in bloodstream of Chagas disease patients. Given that, the comparison of a classical diagnostic method such as blood culture and multiplex quantitative real-time PCR (qPCR) was few explored so far. Therefore, this study aimed to compare the detection and quantification of T. cruzi load in the circulating blood of patients with chronic Chagas disease, using blood culture and qPCR techniques. Methods/Principal findings The multiplex real-time quantitative PCR assay (qPCR) based on TaqMan technology was evaluated in 135 blood samples from 91 patients with chronic Chagas disease presenting indeterminate (asymptomatic, n = 23) and cardiac (chronic cardiomyopathy, n = 68) forms, in comparison with the classical blood culture (BC) technique. The total positivity of qPCR and BC was 58.5% and 49.6%, respectively. The median parasite load of all positive patients was 1.18 [0.39-4.23] par. eq./mL, ranging from 0.01 to 116.10 par. eq./mL. We did not find significant differences between T. cruzi load with age and distinct clinical manifestations of patients. Conclusions/Significance Our data suggest that qPCR can be an auxiliary tool for studies that require T. cruzi isolation from the bloodstream of patients with chronic Chagas disease, after the establishment of a parasite load cut-off that guarantees a relative success rate of parasite isolation using BC technique.
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页数:16
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