Prospective multicenter evaluation of real time PCR Kit prototype for early diagnosis of congenital Chagas disease.


Journal

EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039

Informations de publication

Date de publication:
Jul 2021
Historique:
received: 07 10 2019
revised: 29 05 2021
accepted: 04 06 2021
pubmed: 30 6 2021
medline: 6 8 2021
entrez: 29 6 2021
Statut: ppublish

Résumé

Current algorithm for Congenital Chagas Disease (cCD) diagnosis is unsatisfactory due to low sensitivity of the parasitological methods. Moreover, loss to follow-up precludes final serodiagnosis after nine months of life in many cases. A duplex TaqMan qPCR kit for Trypanosoma cruzi DNA amplification was prospectively evaluated in umbilical cord (UCB) and peripheral venous blood (PVB) of infants born to CD mothers at endemic and non-endemic sites of Argentina. We enrolled and followed-up 370 infants; qPCR was compared to gold-standard cCD diagnosis following studies of diagnostic accuracy guidelines. Fourteen infants (3·78%) had cCD. The qPCR sensitivity and specificity were higher in PVB (72·73%, 99·15% respectively) than in UCB (66·67%, 96·3%). Positive and negative predictive values were 80 and 98·73% and 50 and 98·11% for PVB and UCB, respectively. The Areas under the Curve (AUC) of ROC analysis for qPCR and micromethod (MM) were 0·81 and 0·67 in UCB and 0·86 and 0·68 in PVB, respectively. Parasitic loads ranged from 37·5 to 23,709 parasite equivalents/mL. Discrete typing Unit Tc V was identified in five cCD patients and in six other cCD cases no distinction among Tc II, Tc V or Tc VI was achieved. This first prospective field study demonstrated that qPCR was more sensitive than MM for early cCD detection and more accurate in PVB than in UCB. Its use, as an auxiliary diagnostic tool to MM will provide more accurate records on cCD incidence. FITS SALUD 001-CHAGAS (FONARSEC, MINCyT, Argentina) to the Public-Private Consortium (INGEBI-CONICET, INP-ANLIS MALBRAN and Wiener Laboratories); ERANET-LAC-HD 328 to AGS and PICT 2015-0074 (FONCYT, MinCyT) to AGS and FA.

Sections du résumé

BACKGROUND BACKGROUND
Current algorithm for Congenital Chagas Disease (cCD) diagnosis is unsatisfactory due to low sensitivity of the parasitological methods. Moreover, loss to follow-up precludes final serodiagnosis after nine months of life in many cases. A duplex TaqMan qPCR kit for Trypanosoma cruzi DNA amplification was prospectively evaluated in umbilical cord (UCB) and peripheral venous blood (PVB) of infants born to CD mothers at endemic and non-endemic sites of Argentina.
METHODS METHODS
We enrolled and followed-up 370 infants; qPCR was compared to gold-standard cCD diagnosis following studies of diagnostic accuracy guidelines.
FINDINGS RESULTS
Fourteen infants (3·78%) had cCD. The qPCR sensitivity and specificity were higher in PVB (72·73%, 99·15% respectively) than in UCB (66·67%, 96·3%). Positive and negative predictive values were 80 and 98·73% and 50 and 98·11% for PVB and UCB, respectively. The Areas under the Curve (AUC) of ROC analysis for qPCR and micromethod (MM) were 0·81 and 0·67 in UCB and 0·86 and 0·68 in PVB, respectively. Parasitic loads ranged from 37·5 to 23,709 parasite equivalents/mL. Discrete typing Unit Tc V was identified in five cCD patients and in six other cCD cases no distinction among Tc II, Tc V or Tc VI was achieved.
INTERPRETATION CONCLUSIONS
This first prospective field study demonstrated that qPCR was more sensitive than MM for early cCD detection and more accurate in PVB than in UCB. Its use, as an auxiliary diagnostic tool to MM will provide more accurate records on cCD incidence.
FUNDING BACKGROUND
FITS SALUD 001-CHAGAS (FONARSEC, MINCyT, Argentina) to the Public-Private Consortium (INGEBI-CONICET, INP-ANLIS MALBRAN and Wiener Laboratories); ERANET-LAC-HD 328 to AGS and PICT 2015-0074 (FONCYT, MinCyT) to AGS and FA.

Identifiants

pubmed: 34186488
pii: S2352-3964(21)00243-7
doi: 10.1016/j.ebiom.2021.103450
pmc: PMC8243352
pii:
doi:

Substances chimiques

Reagent Kits, Diagnostic 0

Types de publication

Evaluation Study Journal Article Multicenter Study

Langues

eng

Sous-ensembles de citation

IM

Pagination

103450

Informations de copyright

Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Competing Interest None of the authors received payment or service from a third part at any time. None of the authors have any patents relevant to the work.

Auteurs

Alejandro Francisco Benatar (AF)

Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh) Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr Héctor Torres" (INGEBI) - Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET; Ciudad Autónoma de Buenos Aires, Argentina.

Emmaría Danesi (E)

Centro Nacional de Diagnóstico e Investigación en Endemoepidemias (ANLIS Dr. C. G. Malbrán, Ciudad Autónoma de Buenos Aires, Argentina).

Susana Alicia Besuschio (SA)

Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh) Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr Héctor Torres" (INGEBI) - Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET; Ciudad Autónoma de Buenos Aires, Argentina.

Santiago Bortolotti (S)

Centro de Investigación y Biotecnología (CIBIO) de Wiener Laboratorios SAIC, Rosario, Pcia. de Santa Fe, Argentina.

María Luisa Cafferata (ML)

Departamento en Salud de la Madre y el Niño, Instituto de Efectividad Clínica y Sanitaria, Ciudad Autónoma de Buenos Aires, Argentina.

Juan Carlos Ramirez (JC)

Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh) Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr Héctor Torres" (INGEBI) - Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET; Ciudad Autónoma de Buenos Aires, Argentina.

Constanza Lopez Albizu (CL)

Instituto Nacional de Parasitología Dr. M. Fatala Chabén; Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán); Ciudad Autónoma de Buenos Aires, Argentina.

Karenina Scollo (K)

Instituto Nacional de Parasitología Dr. M. Fatala Chabén; Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán); Ciudad Autónoma de Buenos Aires, Argentina.

María Baleani (M)

Centro de Investigación y Biotecnología (CIBIO) de Wiener Laboratorios SAIC, Rosario, Pcia. de Santa Fe, Argentina.

Laura Lara (L)

Instituto de Maternidad y Ginecología "Nuestra Señora de las Mercedes", San Miguel de Tucumán, Pcia. de Tucumán, Argentina.

Gustavo Agolti (G)

Hospital "Dr. Julio César Perrando", Resistencia, Pcia. de Chaco, Argentina.

Sandra Seu (S)

Hospital Regional "Dr. Ramón Carrillo", Santiago del Estero, Pcia. de Santiago del Estero, Argentina.

Elsa Adamo (E)

Centro Integral de Salud La Banda, La Banda, Pcia. de Santiago del Estero, Argentina.

Raúl Horacio Lucero (RH)

Instituto de Medicina Regional, Universidad Nacional del Nordeste (IMR-UNNE) Resistencia, Provincia de Chaco, Argentina.

Lucía Irazu (L)

Departamento de Calidad. Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán).

Marcelo Rodriguez (M)

Departamento de Calidad. Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán).

Andrés Poeylaut-Palena (A)

Centro de Investigación y Biotecnología (CIBIO) de Wiener Laboratorios SAIC, Rosario, Pcia. de Santa Fe, Argentina.

Silvia Andrea Longhi (SA)

Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh) Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr Héctor Torres" (INGEBI) - Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET; Ciudad Autónoma de Buenos Aires, Argentina.

Mónica Esteva (M)

Instituto Nacional de Parasitología Dr. M. Fatala Chabén; Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán); Ciudad Autónoma de Buenos Aires, Argentina.

Fernando Althabe (F)

Departamento en Salud de la Madre y el Niño, Instituto de Efectividad Clínica y Sanitaria, Ciudad Autónoma de Buenos Aires, Argentina.

Federico Rojkin (F)

Centro de Investigación y Biotecnología (CIBIO) de Wiener Laboratorios SAIC, Rosario, Pcia. de Santa Fe, Argentina.

Jacqueline Bua (J)

Instituto Nacional de Parasitología Dr. M. Fatala Chabén; Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán); Ciudad Autónoma de Buenos Aires, Argentina.

Sergio Sosa-Estani (S)

Departamento en Salud de la Madre y el Niño, Instituto de Efectividad Clínica y Sanitaria, Ciudad Autónoma de Buenos Aires, Argentina; Instituto Nacional de Parasitología Dr. M. Fatala Chabén; Administración Nacional de Laboratorios e Institutos de Salud Carlos G. Malbrán (ANLIS-Malbrán); Ciudad Autónoma de Buenos Aires, Argentina.

Alejandro Gabriel Schijman (AG)

Laboratorio de Biología Molecular de la Enfermedad de Chagas (LaBMECh) Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr Héctor Torres" (INGEBI) - Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET; Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: schijman@dna.uba.ar.

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