Real-Time PCR in HIV/Trypanosoma cruzi Coinfection with and without Chagas Disease Reactivation: Association with HIV Viral Load and CD4+ Level

Abstract

BackgroundReactivation of chronic Chagas disease, which occurs in approximately 20% of patients coinfected with HIV/Trypanosoma cruzi (T. cruzi), is commonly characterized by severe meningoencephalitis and myocarditis. The use of quantitative molecular tests to monitor Chagas disease reactivation was analyzed.MethodologyPolymerase chain reaction (PCR) of kDNA sequences, competitive (C-) PCR and real-time quantitative (q) PCR were compared with blood cultures and xenodiagnosis in samples from 91 patients (57 patients with chronic Chagas disease and 34 with HIV/T. cruzi coinfection), of whom 5 had reactivation of Chagas disease and 29 did not.Principal FindingsqRT-PCR showed significant differences between groups; the highest parasitemia was observed in patients infected with HIV/T. cruzi with Chagas disease reactivation (median 1428.90 T. cruzi/mL), followed by patients with HIV/T. cruzi infection without reactivation (median 1.57 T. cruzi/mL) and patients with Chagas disease without HIV (median 0.00 T. cruzi/mL). Spearman's correlation coefficient showed that xenodiagnosis was correlated with blood culture, C-PCR and qRT-PCR. A stronger Spearman correlation index was found between C-PCR and qRT-PCR, the number of parasites and the HIV viral load, expressed as the number of CD4+ cells or the CD4+/CD8+ ratio.ConclusionsqRT-PCR distinguished the groups of HIV/T. cruzi coinfected patients with and without reactivation. Therefore, this new method of qRT-PCR is proposed as a tool for prospective studies to analyze the importance of parasitemia (persistent and/or increased) as a criterion for recommending pre-emptive therapy in patients with chronic Chagas disease with HIV infection or immunosuppression. As seen in this study, an increase in HIV viral load and decreases in the number of CD4+ cells/mm3 and the CD4+/CD8+ ratio were identified as cofactors for increased parasitemia that can be used to target the introduction of early, pre-emptive therapy.

2 nd Brazilian Consensus on Chagas Disease, 2015

“…A few of cases have been reported of kidney transplant recipients without Chagas disease who were treated with antiparasitic prophylaxis for 14 days, as the organ donors had Chagas disease (308) . In another work, prophylaxis nd Brazilian Consensus on Chagas Disease, 2015 was provided for 60 days to recipients (without Chagas disease) who received livers from donors with Chagas disease (306) as usual strategy; seroconversion was not observed in the surviving recipients, as this may not occur in about 20% of patients in the posttransplant period .…”

Host stress physiology and Trypanosoma haemoparasite infection influence innate immunity in the woylie ( Bettongia penicillata )

“…While Trypanosoma infection status was not found to be affected by FCM or phagocytosis parameters, the degree of parasitaemia or pathology could be affected. Future research could consider quantitatively assessing degree of parasitaemia by, for example, using concentration techniques [58] or real-time quantitative PCR [59]. We also found no association between the presence or absence of ticks and trypanosomes despite suggestions that ticks act as vectors for trypanosomes in woylie [16].…”

Enfermedad de Chagas del sistema nervioso central en un paciente con SIDA demostrada por métodos cuantitativos moleculares

“…En años más recientes se ha desarrollado el diagnóstico cuantitativo por métodos moleculares como el aplicado en nuestro paciente 29 . En efecto, los pacientes con reactivación de enfermedad de Chagas e infección por VIH presentan cargas parasitarias significativamente más altas que los pacientes con ambas condiciones pero sin reactivación (medianas de 1.428 copias/mL versus 1,57 copias/ mL; respectivamente) 29 . Así, las 1.400 copias/mL de T. cruzi detectadas en nuestro paciente en el compartimento sanguíneo están en concordancia con estos estudios.…”