INVESTIGADORES
SCHIJMAN Alejandro Gabriel
artículos
Título:
Analytical Performance of a Multiplex Real-Time PCR Assay Using TaqMan Probes for Quantification of Trypanosoma cruzi Satellite DNA in Blood Samples
Autor/es:
TOMAS DUFFY ; CAROLINA I. CURA ; JUAN C. RAMIREZ . ; TERESA ABATE ; NELLY M. CAYO ; RUDY PARRADO ; ZORAIDA DIAZ BELLO ; ELSA VELAZQUEZ ; ARTURO MUÑOZ-CALDERON; NATALIA A. JUIZ ; JOAQUıN BASILE ; LINETH GARCIA; ADELINA RIARTE ; JULIO NASSER; SUSANA B. OCAMPO ; ZAIDA E. YADON ; FAUSTINO TORRICO; BELKISYOLE ALARCON DE NOYA ; ISABELA RIBEIRO; ALEJANDO GABRIEL SCHIJMAN
Revista:
PLOS NEGLECTED TROPICAL DISEASES
Editorial:
PUBLIC LIBRARY SCIENCE
Referencias:
Lugar: San Francisco; Año: 2013 vol. 7 p. 1 - 11
ISSN:
1935-2735
Resumen:
Abstract
Background:
The analytical validation of sensitive, accurate and standardized Real-Time PCR methods for
Trypanosoma cruzi
quantification is crucial to provide a reliable laboratory tool for diagnosis of recent infections as well as for monitoring
treatment efficacy.
Methods/Principal Findings:
We have standardized and validated a multiplex Real-Time quantitative PCR assay (qPCR)
based on TaqMan technology, aiming to quantify
T. cruzi
satellite DNA as well as an internal amplification control (IAC) in a
single-tube reaction. IAC amplification allows rule out false negative PCR results due to inhibitory substances or loss of DNA
during sample processing. The assay has a limit of detection (LOD) of 0.70 parasite equivalents/mL and a limit of
quantification (LOQ) of 1.53 parasite equivalents/mL starting from non-boiled Guanidine EDTA blood spiked with
T. cruzi
CL-
Brener stock. The method was evaluated with blood samples collected from Chagas disease patients experiencing different
clinical stages and epidemiological scenarios: 1- Sixteen Venezuelan patients from an outbreak of oral transmission, 2- Sixty
three Bolivian patients suffering chronic Chagas disease, 3- Thirty four Argentinean cases with chronic Chagas disease,
Twenty seven newborns to seropositive mothers, 5- A seronegative receptor who got infected after transplantation with a
cadaveric kidney explanted from an infected subject.
Conclusions/Significance:
The performing parameters of this assay encourage its application to early assessment of
T. cruzi
infection in cases in which serological methods are not informative, such as recent infections by oral contamination or
congenital transmission or after transplantation with organs from seropositive donors, as well as for monitoring Chagasdisease patients under etiological treatment