The Trypanosoma cruzi Satellite DNA OligoC-TesT and Trypanosoma cruzi Kinetoplast DNA OligoC-TesT for Diagnosis of Chagas Disease: A Multi-cohort Comparative Evaluation Study

Abstract

BackgroundThe Trypanosoma cruzi satellite DNA (satDNA) OligoC-TesT is a standardised PCR format for diagnosis of Chagas disease. The sensitivity of the test is lower for discrete typing unit (DTU) TcI than for TcII-VI and the test has not been evaluated in chronic Chagas disease patients.Methodology/Principal FindingsWe developed a new prototype of the OligoC-TesT based on kinetoplast DNA (kDNA) detection. We evaluated the satDNA and kDNA OligoC-TesTs in a multi-cohort study with 187 chronic Chagas patients and 88 healthy endemic controls recruited in Argentina, Chile and Spain and 26 diseased non-endemic controls from D.R. Congo and Sudan. All specimens were tested in duplicate. The overall specificity in the controls was 99.1% (95% CI 95.2%–99.8%) for the satDNA OligoC-TesT and 97.4% (95% CI 92.6%–99.1%) for the kDNA OligoC-TesT. The overall sensitivity in the patients was 67.9% (95% CI 60.9%–74.2%) for the satDNA OligoC-TesT and 79.1% (95% CI 72.8%–84.4%) for the kDNA OligoC-Test.Conclusions/SignificanceSpecificities of the two T. cruzi OligoC-TesT prototypes are high on non-endemic and endemic controls. Sensitivities are moderate but significantly (p = 0.0004) higher for the kDNA OligoC-TesT compared to the satDNA OligoC-TesT.

Epidemiology of Trypanosoma evansi and Trypanosoma vivax in domestic animals from selected districts of Tigray and Afar regions, Northern Ethiopia

“…Plasma was stored at 4°C until testing for specific antibodies with CATT/ T.evansi and subsequently frozen at −20°C. From the remaining blood specimen, 500 μL of buffy coat layer were collected by means of a micropipette with a filter tip and mixed with an equal volume of guanidium EDTA buffer (GEB; 6 M guanidium chloride, 0.2 M EDTA, pH 8.0) and stored at ambient temperature until DNA extraction [ 68 ]. Of those animals that were parasitologically positive, part of the buffy coat was collected for cryopreservation in liquid nitrogen for later isolation of the parasite according to Pyana et al [ 69 ].…”

Commercial enzyme-linked immunosorbent assay versuspolymerase chain reaction for the diagnosis of chronic Chagas disease: a systematic review and meta-analysis

“…Finally, data concerning PCR were extracted from 24 reports ( Avila et al 1993 , Wincker et al 1994 , 1997 , Britto et al 1995 , Espinoza et al 1996 , Junqueira et al 1996 , Carriazo et al. 1998 , Chiaramonte et al 1999 , Gomes et al 1999 , Ribeiro-dos-Santos et al 1999 , Castro et al 2002 , Gutierrez et al 2004 , Duarte et al 2006 , Gil et al 2007 , Piron et al 2007 , Fitzwater et al 2008 , Deborggraeve et al 2009 , Ferrer et al 2009, 20 , 2013 , Ramírez et al 2009 , Batista et al 2010 , Gilber et al 2013 , Sabino et al 2013) , but only one commercial test was found ( Deborggraeve et al 2009 , De Winne et al 2014) . However, the original authors modified the commercial version in two of the studies for research purposes (De Winne et al 2014) .…”

Mast Cell Coupling to the Kallikrein–Kinin System Fuels Intracardiac Parasitism and Worsens Heart Pathology in Experimental Chagas Disease

“…The standards for the qPCRs were generated using DNA from T. cruzi (Dm28c) epimastigotes. Considering that one parasite contains about 100 fg of DNA ( 46 ), we designed a curve with initial 2.5 ng (equivalent to 25,000 parasites with C t -mean 19) and serially diluted (1:10) until the lower limit of 2.5 fg (0.025 parasites with a C t -mean 37). The reactions were performed in a StepOne sequence detection system (Applied Biosystems, Warrington, UK) using SYBR Green PCR Master Mix (Applied Biosystems, Warrington, UK).…”