12 research outputs found
Prospective multicenter evaluation of real time PCR Kit prototype for early diagnosis of congenital Chagas disease
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: We enrolled and followed-up 370 infants; qPCR was compared to gold-standard cCD diagnosis following studies of diagnostic accuracy guidelines. Findings: 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: 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: 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.Fil: Benatar, Alejandro Francisco. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Danesi, EmmarĂa. DirecciĂłn Nacional de Instituto de InvestigaciĂłn.AdministraciĂłn Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"; ArgentinaFil: Besuschio, Susana Alicia. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Bortolotti, Santiago. Wiener Laboratorios SAIC; ArgentinaFil: Cafferata, MarĂa Luisa. Instituto de Efectividad ClĂnica y Sanitaria; ArgentinaFil: Ramirez Gomez, Juan Carlos. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Lopez Albizu, Maria Constanza. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de ParasitologĂa "Dr. Mario Fatala Chaben”; ArgentinaFil: Scollo, Karenina. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de ParasitologĂa "Dr. Mario Fatala Chaben”; ArgentinaFil: Baleani, MarĂa. Wiener Laboratorios SAIC; ArgentinaFil: Lara, Laura. Instituto de Maternidad y GinecologĂa ''Nuestra Señora de las Mercedes''; ArgentinaFil: Agolti, Gustavo. Gobierno de la Provincia de Chaco. Hospital Julio CĂ©sar Perrando; ArgentinaFil: Seu, Sandra. Gobierno de la Provincia de Santiago del Estero. Hospital Regional Dr. RamĂłn Carrillo; ArgentinaFil: Adamo, Elsa. Provincia de Santiago del Estero. Centro Integral de Salud La Banda; ArgentinaFil: Lucero, Raul Horacio. Universidad Nacional del Nordeste. Instituto de Medicina Regional; ArgentinaFil: Irazu, LucĂa. DirecciĂłn Nacional de Instituto de InvestigaciĂłn.AdministraciĂłn Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"; ArgentinaFil: Rodriguez, Marcelo. DirecciĂłn Nacional de Instituto de InvestigaciĂłn.AdministraciĂłn Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"; ArgentinaFil: Poeylaut Palena, AndrĂ©s Alberto. Wiener Laboratorios SAIC; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas; ArgentinaFil: Longhi, Silvia Andrea. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Esteva, MĂłnica InĂ©s. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de ParasitologĂa "Dr. Mario Fatala Chaben”; ArgentinaFil: Althabe, Fernando. Instituto de Efectividad ClĂnica y Sanitaria; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas; ArgentinaFil: Rojkin, Federico. Wiener Laboratorios SAIC; ArgentinaFil: Bua, Jacqueline Elena. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de ParasitologĂa "Dr. Mario Fatala Chaben”; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas; ArgentinaFil: Sosa-Estani, Sergio Alejandro. Instituto de Efectividad ClĂnica y Sanitaria; Argentina. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de ParasitologĂa "Dr. Mario Fatala Chaben”; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas; ArgentinaFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Instituto de Investigaciones en IngenierĂa GenĂ©tica y BiologĂa Molecular "Dr. HĂ©ctor N. Torres"; ArgentinaFil: Congenital Chagas Disease Study Group. No especifĂca
Congenital bone marrow failure in DNA-PKcs mutant mice associated with deficiencies in DNA repair
The nonhomologous end-joining (NHEJ) pathway is essential for radioresistance and lymphocyte-specific V(D)J (variable [diversity] joining) recombination. Defects in NHEJ also impair hematopoietic stem cell (HSC) activity with age but do not affect the initial establishment of HSC reserves. In this paper, we report that, in contrast to deoxyribonucleic acid (DNA)–dependent protein kinase catalytic subunit (DNA-PKcs)–null mice, knockin mice with the DNA-PKcs(3A/3A) allele, which codes for three alanine substitutions at the mouse Thr2605 phosphorylation cluster, die prematurely because of congenital bone marrow failure. Impaired proliferation of DNA-PKcs(3A/3A) HSCs is caused by excessive DNA damage and p53-dependent apoptosis. In addition, increased apoptosis in the intestinal crypt and epidermal hyperpigmentation indicate the presence of elevated genotoxic stress and p53 activation. Analysis of embryonic fibroblasts further reveals that DNA-PKcs(3A/3A) cells are hypersensitive to DNA cross-linking agents and are defective in both homologous recombination and the Fanconi anemia DNA damage response pathways. We conclude that phosphorylation of DNA-PKcs is essential for the normal activation of multiple DNA repair pathways, which in turn is critical for the maintenance of diverse populations of tissue stem cells in mice
Comorbidity and high viral load linked to clinical presentation of respiratory human bocavirus infection
Human bocavirus (HBoV) is a new parvovirus associated with acute respiratory tract infection (ARTI). In order to evaluate HBoV significance as an agent of acute respiratory disease, we screened 1,135 respiratory samples from children and adults with and without symptoms during two complete calendar years. HBoV1 prevalence in patients with ARTI was 6.33 % in 2011 and 11.64 % in 2012, including neonatal and adult patients. HBoV1 was also detected in 3.77 % of asymptomatic individuals. The co-detection rate was 78.1 %. Among children, 87 % were clinically diagnosed with lower respiratory infection (no significant differences between patients with and without coinfection), and 31 % exhibited comorbidities. Pediatric patients with comorbidities were significantly older than patients without comorbidities. Patients with ARTI had either high or low viral load, while controls had only low viral load, but there were no clinical differences between patients with high or low viral load. In conclusion, we present evidence of the pathogenic potential of HBoV1 in young children with ARTI. Since patients with HBoV1-single infection are not significantly different from those with coinfection with respect to clinical features, the virus can be as pathogenic by itself as other respiratory agents are. Furthermore, an association between high HBoV1 load and disease could not be demonstrated in this study, but all asymptomatic individuals had low viral loads. Also, children with comorbidities are susceptible to HBoV1 infection at older ages than previously healthy children. Thus, the clinical presentation of infection may occur depending on both viral load and the particular interaction between the HBoV1 and the host.Fil: Ghietto, LucĂa MarĂa. Universidad Nacional de CĂłrdoba. Facultad de Medicina. Instituto de VirologĂa ; ArgentinaFil: Majul, Diego. Universidad Nacional de Cordoba. Hospital de Niños; ArgentinaFil: Ferreyra Soaje, Patricia. Universidad Nacional de Cordoba. Hospital de Niños; ArgentinaFil: Baumeister, Elsa. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorios e Instituto de Salud ; ArgentinaFil: Avaro, MartĂn. DirecciĂłn Nacional de Instituto de InvestigaciĂłn. AdministraciĂłn Nacional de Laboratorios e Instituto de Salud ; ArgentinaFil: Insfran, Constanza. Universidad Nacional de CĂłrdoba. Facultad de Medicina. Instituto de VirologĂa ; ArgentinaFil: Mosca, Liliana. Universidad Nacional de Cordoba. Hospital de Niños; ArgentinaFil: Cámara, Alicia. Universidad Nacional de CĂłrdoba. Facultad de Medicina. Instituto de VirologĂa ; ArgentinaFil: Moreno, Laura Beatriz. Universidad Nacional de Cordoba. Hospital de Niños; ArgentinaFil: Adamo, Maria Pilar. Universidad Nacional de CĂłrdoba. Facultad de Medicina. Instituto de VirologĂa ; Argentin