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- Title
Trypanosoma cruzi loop-mediated isothermal amplification (Trypanosoma cruzi Loopamp) kit for detection of congenital, acute and Chagas disease reactivation.
- Authors
Besuschio, Susana A.; Picado, Albert; Muñoz-Calderón, Arturo; Wehrendt, Diana P; Fernández, Marisa; Benatar, Alejandro; Diaz-Bello, Zoraida; Irurtia, Cecilia; Cruz, Israel; Ndung'u, Joseph M; Cafferata, María L; Montenegro, Graciela; Sosa Estani, Sergio; Lucero, Raúl H.; Alarcón de Noya, Belkisyole; Longhi, Silvia A; Schijman, Alejandro G
- Abstract
A Trypanosoma cruzi Loopamp kit was recently developed as a ready-to-use diagnostic method requiring minimal laboratory facilities. We evaluated its diagnostic accuracy for detection of acute Chagas disease (CD) in different epidemiological and clinical scenarios. In this retrospective study, a convenience series of clinical samples (venous blood treated with EDTA or different stabilizer agents, heel-prick blood in filter paper or cerebrospinal fluid samples (CSF)) from 30 infants born to seropositive mothers (13 with congenital CD and 17 noninfected), four recipients of organs from CD donors, six orally–infected cases after consumption of contaminated guava juice and six CD patients coinfected with HIV at risk of CD reactivation (N = 46 patients, 46 blood samples and 1 CSF sample) were tested by T. cruzi Loopamp kit (Tc LAMP) and standardized quantitative real-time PCR (qPCR). T. cruzi Loopamp accuracy was estimated using the case definition in the different groups as a reference. Cohen's kappa coefficient (κ) was applied to measure the agreement between Tc LAMP (index test) and qPCR (reference test). Sensitivity and specificity of T. cruzi Loopamp kit in blood samples from the pooled clinical groups was 93% (95% CI: 77–99) and 100% (95% CI: 80–100) respectively. The agreement between Tc LAMP and qPCR was almost perfect (κ = 0.92, 95% CI: 0.62–1.00). The T. cruzi Loopamp kit was sensitive and specific for detection of T. cruzi infection. It was carried out from DNA extracted from peripheral blood samples (via frozen EDTA blood, guanidine hydrochloride-EDTA blood, DNAgard blood and dried blood spots), as well as in CSF specimens infected with TcI or TcII/V/VI parasite populations. The T. cruzi Loopamp kit appears potentially useful for rapid detection of T. cruzi infection in congenital, acute and CD reactivation due to HIV infection. Author summary: The aim of this study was to evaluate a kit prototype (T. cruzi Loopamp or Tc LAMP) based on loop-mediated isothermal amplification (LAMP) for molecular diagnosis of acute Chagas disease in well-characterized individuals, collected in real life conditions, such as newborns or infants born to Chagas disease mothers aiming to detect congenital Chagas disease, recipients of organs from Chagas disease donors who acquired T. cruzi infection after transplantation, persons with oral Chagas disease acquired after consumption of a contaminated meal, and HIV/T. cruzi coinfected patients at risk of Chagas disease reactivation due to immunosuppression. Different types of clinical samples were tested; peripheral blood samples treated with ethylenediamine tetraacetic acid (EDTA) as anticoagulant or with guanidine hydrochloride or DNAgard stabilizer agents, as well as dried blood spots and cerebrospinal fluid samples. The performance of Tc LAMP was estimated using the case definition in the different groups as a reference and all test kit results were compared to those obtained using standardized duplex real-time PCR (qPCR) in the same clinical samples. Sensitivity and specificity of Tc LAMP in blood samples from the pooled clinical groups were 93% (95% CI: 77–99) and 100% (95% CI: 80–100) respectively. The agreement between Tc LAMP and qPCR was almost perfect (κ index of Cohen = 0.92, 95% CI: 0.62–1.00). Tc LAMP was sensitive and specific for detection of T. cruzi infection in the tested samples, encouraging prospective field studies to validate its use for rapid detection of T. cruzi infection in congenital, acute and CD reactivation due to HIV infection.
- Subjects
CHAGAS' disease; TRYPANOSOMA cruzi; ACUTE diseases; NEWBORN infants; CONGENITAL disorders; CONGENITAL hypothyroidism
- Publication
PLoS Neglected Tropical Diseases, 2020, Vol 14, Issue 8, p1
- ISSN
1935-2727
- Publication type
Article
- DOI
10.1371/journal.pntd.0008402