Trypanosoma cruzi loop-mediated isothermal amplification (Trypanosoma cruzi Loopamp) kit for detection of congenital, acute and Chagas disease reactivation

Autor: Cecilia Irurtia, Raul Horacio Lucero, Maria Luisa Cafferata, Albert Picado, Belkisyolé Alarcón de Noya, Silvia Andrea Longhi, Alejandro Francisco Benatar, Sergio Sosa Estani, Diana Patricia Wehrendt, Susana Alicia Besuschio, Israel Cruz, Alejandro G. Schijman, Graciela Montenegro, Joseph Mathu Ndung'u, Zoraida Díaz-Bello, Arturo Muñoz-Calderón, Marisa Liliana Fernandez
Přispěvatelé: KfW Development Bank, Ministerio de Ciencia, Tecnología e Innovación (Argentina), Latin America and the Caribbean Countries (LAC), Unión Europea, KfW Entwicklungs bank, Argentinean Ministry of Science Technology and Innovation, Network of the European Union (EU), European Union
Rok vydání: 2020
Předmět:
Male
0301 basic medicine
Chagas disease
Physiology
RC955-962
HIV Infections
Artificial Gene Amplification and Extension
Nervous System
Polymerase Chain Reaction
Guanidines
law.invention
purl.org/becyt/ford/1 [https]
Medical Conditions
0302 clinical medicine
Cerebrospinal fluid
law
Arctic medicine. Tropical medicine
Medicine and Health Sciences
Medicine
Parasite hosting
DNA extraction
Polymerase chain reaction
Cerebrospinal Fluid
Protozoans
biology
Coinfection
Eukaryota
Venous blood
ACUTE INFECTIONS
Body Fluids
Chemistry
Blood
Infectious Diseases
Physical Sciences
Female
Molecular diagnosis
Public aspects of medicine
RA1-1270
Anatomy
Nucleic Acid Amplification Techniques
Research Article
Neglected Tropical Diseases
Trypanosoma
Trypanosoma cruzi
030231 tropical medicine
Loop-mediated isothermal amplification
Real-Time Polymerase Chain Reaction
Research and Analysis Methods
Sensitivity and Specificity
03 medical and health sciences
Extraction techniques
LAMP
Parasitic Diseases
Humans
Chagas Disease
Molecular Biology Techniques
purl.org/becyt/ford/1.6 [https]
Molecular Biology
Retrospective Studies
Protozoan Infections
business.industry
Infant
Newborn
Organisms
Chemical Compounds
Public Health
Environmental and Occupational Health
Infant
Biology and Life Sciences
DNA
Protozoan
Tropical Diseases
biology.organism_classification
medicine.disease
Virology
Transplant Recipients
Parasitic Protozoans
030104 developmental biology
business
Zdroj: CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
PLoS Neglected Tropical Diseases
Repisalud
Instituto de Salud Carlos III (ISCIII)
PLoS Neglected Tropical Diseases, Vol 14, Iss 8, p e0008402 (2020)
ISSN: 1935-2735
Popis: 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.
Databáze: OpenAIRE
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