Search

We report a patient with lobomycosis caused by Paracoccidioides loboi fungi in the Andes-Amazon region of Bolivia. We examined clinical, epidemiologic, and phylogenetic data and describe potential transmission/environmental aspects of infection. Continued surveillance and identification of lobomycosis cases in South America are crucial to prevent the spread of this disease.

We report a patient from Panama who had lobomycosis caused by Paracoccidioides (Lacazia) loboi. We used combined clinical-epidemiologic and phylogenetic data, including a new gene sequence dataset on this fungus in Panama, for analysis. Findings contribute useful insights to limited knowledge of this fungal infection in the Mesoamerican Biologic Corridor.

Abstract. Background:. Epidemiological studies in patients with neuropathic pain demonstrate a strong association with psychiatric conditions such as anxiety; however, the precipitating pathology between these symptoms remains unclear. To investigate this, we studied the effects of lifelong stress on levels of neuropathic pain–like behavior and conversely, the effects of chronic neuropathic injury on anxiety-like status in male and female mice. In addition, we assayed this link in painful and painless diabetic peripheral neuropathy patients. Methods:. Male and female mice were subject to ongoing life-stress or control living conditions. Baseline sensitivity and anxiety tests were measured followed by spared nerve injury (SNI) to the sciatic nerve. Subsequent sensory testing occurred until 3 weeks after SNI followed by anxiety tests between 4 and 6 weeks after SNI. Results:. Levels of tactile or cold allodynia did not differ between adult mice subject to lifelong chronic stress, relative to nonstressed controls, for at least 3 weeks after SNI. By contrast, longer-term neuropathic mice of both sexes displayed pronounced anxiety-like behavior, regardless of exposure to stress. If sex differences were present, females usually exhibited more pronounced anxiety-like behavior. These ongoing anxiety behaviors were corroborated with plasma corticosterone levels in distinct animal groups. In addition, data from patients with painful and nonpainful diabetic neuropathy showed a clear relationship between ongoing pain and anxiety, with females generally more affected than males. Discussion:. Taken together, these data demonstrate a strong link between chronic neuropathic pain and chronic anxiety, with the driver of this comorbidity being neuropathic pain as opposed to on-going stress.

Abstract Background Bacteria of the genus Borrelia are agents of disease in both domestic animals and humans and pose a significant public health risk. Borrelia species have complex transmission cycles, often using rodents as vertebrate reservoir hosts. These bacteria are classified into three well-defined monophyletic groups: Borrelia burgdorferi sensu lato (Bbsl) complex, the relapsing fever (RF) group, and a third group associated with reptiles and echidnas. Moreover, a new group of Borrelia associated with rodents has recently been proposed, as these bacteria form a phylogenetic group separated from the previously mentioned groups. This study aimed to investigate the presence of DNA of Borrelia spirochetes in rodents in specific areas of the Colombian Caribbean. Methods A total of 155 rodent spleen samples were selected from the tissue bank. These samples were obtained in the departments of La Guajira and Córdoba (Northern Colombia). DNA extraction and specific real-time polymerase chain reaction (PCR) targeting Borrelia 16S ribosomal RNA (rRNA) gene were performed, followed by nested PCR (nPCR) on positive samples to obtain larger fragments of the 16S rRNA gene and characterize the flaB gene. Alignments of generated sequences and ortholog sequences downloaded from Genbank were performed in Clustal Omega. A phylogenetic tree was built with the maximum likelihood method in IQTREE. Results Spleen samples from rodents of the genera Heteromys, Mus, Necromys, Olygoryzomys, Proechymis, Rattus, Sigmodon, and Zygodontomys were processed. Overall, 6.5% (4/162) of the animals tested positive for Borrelia by real-time PCR. All quantitative PCR (qPCR)-positive samples were also positive for nPCR targeting the 16S rRNA gene, yielding fragments of 344–408 bp and 603–673 bp from two Sigmodon rodents and two Zygodontomys rodents from La Guajira and Córdoba. All samples were negative for the flaB gene. Only samples from Zygodontomys rodents presented good quality sequences. A BLASTn analysis showed a percentage of identity ranging between 98.16 and 96.06% with Borrelia sp. R57. Phylogenetic analysis revealed that sequences of the present study clustered with species of the recently proposed Borrelia “rodent group.” Conclusions This is the first detection of borreliae of the “rodent group” in South America. Our results reaffirm the occurrence of a group of spirochetes associated with rodents, extending its geographic distribution to the Colombian Caribbean. Graphical abstract

La fibrilación auricular es la arritmia cardíaca más frecuente del mundo y, de igual manera, es la arritmia predominante en los servicios de urgencias. Cada vez hay más prevalencia de esta enfermedad y, en consecuencia, mayores probabilidades de que los médicos en los servicios de urgencias se enfrenten a ella. Las estrategias de tratamiento estándar de la fibrilación auricular suelen ser “conservadoras” y tradicionalmente han sido el control de la frecuencia, la anticoagulación y la hospitalización. La principal motivación de este tipo de estrategia es prevenir el accidente cerebrovascular. Cuando pasa el tiempo y se produce remodelado atrial, la fibrilación auricular es cada vez más refractaria a la conversión al ritmo sinusal y cuanto más tiempo persiste y el daño cardíaco estructural aumenta, más se incrementa la duración de la fibrilación auricular. Así, en el servicio de urgencias, el manejo adecuado de un primer episodio o de una complicación, puede minimizar costos y reducir exposiciones innecesarias a terapias inadecuadas. Esta revisión aborda el manejo de la fibrilación auricular en el servicio de urgencias con base en la evidencia que se tiene disponible en el momento.

Introducción: La fibrilación auricular es la taquiarritmia sostenida más frecuente del ser humano y su manejo requiere un abordaje holístico para que los resultados sean óptimos. Objetivo: Describir las características sociodemográficas y clínicas de los pacientes hospitalizados o intervenidos, con diagnóstico de fibrilación auricular, atendidos en una clínica de cuarto nivel de complejidad en Colombia. Método: Se realizó un estudio observacional, retrospectivo, a partir de registros de pacientes hospitalizados o intervenidos con diagnóstico de fibrilación auricular. Se tomó como referencia el diagnóstico de fibrilación auricular (CIE 10 I48X), durante los años 2017 y 2018. Se realizó análisis univariado y bivariado con Excel, SPSS y Epidat. Resultados: El promedio de edad de los pacientes hospitalizados fue de 69.81 años, con ligera predominancia del sexo masculino. La estancia hospitalaria fue de 6.62 en 2017 y de 5.29 días en 2018. Se encontró correlación entre los días de estancia hospitalaria y la edad de los pacientes. La comorbilidad más frecuentes en los pacientes hospitalizados fue hipertensión arterial, cardiopatía isquémica y enfermedad pulmonar obstructiva crónica. El 18.04% no tuvieron comorbilidad. Las muertes intrahospitalarias estuvieron asociadas, con mayor frecuencia, a insuficiencia cardíaca. La cardioversión eléctrica fue el procedimiento electrofisiológico más usado, seguida del aislamiento de venas pulmonares. El porcentaje de complicaciones fue menor en 2018 que en 2017. Conclusiones: Existe menor proporción de comorbilidad cuando se compara esta s

La disyunción del anillo valvular mitral es una anormalidad estructural que consiste en un desplazamiento auricular del punto de articulación de la válvula mitral, lo que lleva a una relación espacial alterada entre la válvula y la pared ventricular posterior adyacente. Los estudios han demostrado una relación entre la disyunción del anillo mitral, las arritmias ventriculares y la degeneración fibrosa miocárdica, lo que incrementa el riesgo de muerte súbita, en especial en mujeres jóvenes. Se presenta el caso de una mujer de 30 años, sin antecedentes relevantes, con palpitaciones frecuentes, en quien se documentó un aumento del automatismo ventricular con incremento progresivo de la carga arrítmica hasta el 20% a pesar del manejo farmacológico. En la resonancia cardiaca se apreció un desplazamiento de 15 mm hacia la aurícula de las valvas de la mitral compatible con disyunción del anillo mitral, además de focos de realce tardío de distribución mesocárdica, no coronariana. Ante los hallazgos y la persistencia de los síntomas, se llevó a estudio electrofisiológico, mapeo 3D y ablación. En la ecografía intracardiaca se apreciaron dos regiones hiperecoicas, en la longitud del músculo papilar anterior y en el mesocardio de la base del músculo papilar posterior; ambos focos relacionados con los sitios morfológicos de interés, en los cuales se aplicó energía de radiofrecuencia. Durante el procedimiento presentó un episodio de fibrilación ventricular que se consideró una extrasistolia ventricular maligna, por lo que se implantó un cardiodesfibrilador para prevención de muerte súbita. Se revisa la literatura y se analizan las relaciones fisiopatológicas existentes entre la disyunción del anillo mitral, los complejos ventriculares prematuros y el riesgo de muerte súbita.

Abstract Background Colombia was the second most affected country during the American Zika virus (ZIKV) epidemic, with over 109,000 reported cases. Despite the scale of the outbreak, limited genomic sequence data were available from Colombia. We sought to sequence additional samples and use genomic epidemiology to describe ZIKV dynamics in Colombia. Methods We sequenced ZIKV genomes directly from clinical diagnostic specimens and infected Aedes aegypti samples selected to cover the temporal and geographic breadth of the Colombian outbreak. We performed phylogeographic analysis of these genomes, along with other publicly-available ZIKV genomes from the Americas, to estimate the frequency and timing of ZIKV introductions to Colombia. Results We attempted PCR amplification on 184 samples; 19 samples amplified sufficiently to perform sequencing. Of these, 8 samples yielded sequences with at least 50% coverage. Our phylogeographic reconstruction indicates two separate introductions of ZIKV to Colombia, one of which was previously unrecognized. We find that ZIKV was first introduced to Colombia in February 2015 (95%CI: Jan 2015 – Apr 2015), corresponding to 5 to 8 months of cryptic ZIKV transmission prior to confirmation in September 2015. Despite the presence of multiple introductions, we find that the majority of Colombian ZIKV diversity descends from a single introduction. We find evidence for movement of ZIKV from Colombia into bordering countries, including Peru, Ecuador, Panama, and Venezuela. Conclusions Similarly to genomic epidemiological studies of ZIKV dynamics in other countries, we find that ZIKV circulated cryptically in Colombia. More accurately dating when ZIKV was circulating refines our definition of the population at risk. Additionally, our finding that the majority of ZIKV transmission within Colombia was attributable to transmission between individuals, rather than repeated travel-related importations, indicates that improved detection and control might have succeeded in limiting the scale of the outbreak within Colombia.

Abstract Background The aim of this study was to evaluate the frequency of toxigenic C. difficile and C. perfringens infections at health care facility-onset (HCFO) and community-onset (CO), in two health care centers (HCC) in Bogotá, Colombia. A total of 220 stool samples from patients presenting diarrhea acquired at HCFO or CO were analyzed by several PCR tests. Results We found that 65.5% (n = 144) of the population had C. difficile infection, followed by toxigenic C. difficile with 57.3% (n = 126), and finally toxigenic C. perfringens with a frequency of 32.7% (n = 72). Conclusions This study is the first molecular detection and characterization of C. difficile and C. perfringens in HCFO and CO in Latin America and demonstrates a relevant frequency of these two species, including coinfection and strikingly diverse toxigenic profiles, especially in the CO.

Venezuela’s tumbling economy and authoritarian rule have precipitated an unprecedented humanitarian crisis. Hyperinflation rates now exceed 45,000%, and Venezuela’s health system is in free fall. The country is experiencing a massive exodus of biomedical scientists and qualified healthcare professionals. Reemergence of arthropod-borne and vaccine-preventable diseases has sparked serious epidemics that also affect neighboring countries. In this article, we discuss the ongoing epidemics of measles and diphtheria in Venezuela and their disproportionate impact on indigenous populations. We also discuss the potential for reemergence of poliomyelitis and conclude that action to halt the spread of vaccine-preventable diseases within Venezuela is a matter of urgency for the country and the region. We further provide specific recommendations for addressing this crisis.

Multilocus sequence typing (MLST) has become a useful tool for studying the genetic diversity of important public health pathogens, such as Chlamydia trachomatis (Ct). Four MLST schemes have been proposed for Ct (data available from Chlamydiales MLST databases). However, the lack of a sole standardized scheme represents the greatest limitation regarding typing this species. This study was thus aimed at evaluating the usefulness of the four MLST schemes available for Ct, describing each molecular marker's pattern and its contribution toward a description of intra-specific genetic diversity and population structure. The markers for each scheme, showed a variable power of dicrimination, exhibiting in some cases over estimation in the determination of Sequence Types (STs). However, individual analysis of each locus's typing efficiency and discrimination power led to identifying 8 markers as having a suitable pattern for intra-specific typing. analyzing the 8 candidate markers gave a combination of 3 of these loci as an optimal scheme for identifying a large amount of STs, maximizing discrimination power whilst maintaining suitable typing efficiency. One scheme was compared against core genome phylogenies, finding a higher typing resolution through the last approach. These results confirm once again that although complete genome data, in particular from core genome MLST (cgMLST) allow a high resolution clustering for Ct isolates. There are combinations of molecular markers that could generate equivalent results, with the advantage of representing an easy implementation strategy and lower costs leading to contribute to the monitoring and molecular epidemiology of Ct.

Members of the Cryptococcus complex, includes Cryptococcus neoformans (most common fungal infection of the brain) and Cryptococcus gattii (high-impact emerging pathogen worldwide). Currently, the fungal multilocus sequence typing database (Fungal MLST Database) constitutes a valuable data repository of the genes used for molecular typing of these pathogens. We analyzed the data available in the Fungal MLST Database for seven housekeeping genes, with the aim to evaluate its contribution in the description of intra-taxa diversity, population genetic structure, and evolutionary patterns. Although the Fungal MLST Database has a greater number of reports for C. neoformans (n = 487) than for C. gattii (n = 344), similar results were obtained for both species in terms of allelic diversity. Phylogenetic reconstructions revealed grouping by molecular type in both species and allowed us to propose differences in evolutionary patterns (gradualism in the case of C. neoformans and punctuated evolution in the case of C. gattii). In addition, C. neoformans showed a population genetic structure consisting of 37 clonal complexes (CCs; CC1 being predominant), high crosslinking [without sequence type (ST) grouping by molecular type], marked divergence events in phylogenetic analysis, and few introgression events (mainly between VNI and VNIV). By contrast, C. gattii showed 50 CCs (with greater homogeneity in ST number by CC) and clustering by molecular type with marked crosslinking events in phylogenetic networks being less evident. Understanding relationships at the molecular level for species of the Cryptococcus complex, based on the sequences of the housekeeping genes, provides information for describing the evolutionary history of these emerging pathogens.

Clostridium difficile (CD) produces antibiotic associated diarrhea and leads to a broad range of diseases. The source of CD infection (CDI) acquisition and toxigenic profile are factors determining the impact of CD. This study aimed at detecting healthcare facility onset- (HCFO) and community-onset (CO) CDI and describing their toxigenic profiles in Bogotá, Colombia. A total of 217 fecal samples from patients suffering diarrhea were simultaneously submitted to two CDI detection strategies: (i) in vitro culture using selective chromogenic medium (SCM; chromID, bioMérieux), followed verification by colony screening (VCS), and (ii) molecular detection targeting constitutive genes, using two conventional PCR tests (conv.PCR) (conv.16S y conv.gdh) and a quantitative test (qPCR.16s). The CD toxigenic profile identified by any molecular test was described using 6 tests independently for describing PaLoc and CdtLoc organization. High overall CDI frequencies were found by both SCM (52.1%) and conv.PCR (45.6% for conv.16S and 42.4% for conv.gdh), compared to reductions of up to half the frequency by VCS (27.2%) or qPCR.16S (22.6%). Infection frequencies were higher for SCM and conv.16S regarding HCFO but greater for CO concerning conv.gdh, such differences being statistically significant. Heterogeneous toxigenic profiles were found, including amplification with lok1/3 primers simultaneously with other PaLoc markers (tcdA, tcdB or tcdC). These findings correspond the first report regarding the differential detection of CDI using in vitro culture and molecular detection tests in Colombia, the circulation of CD having heterogeneous toxigenic profiles and molecular arrays which could affect the impact of CDI epidemiology.

Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America.

Las infecciones por hongos en dispositivos cardiacos son excepcionales y los casos reportados por Aspergillus son aún más escasos. Su diagnóstico requiere de un alto índice de sospecha, además de la extracción completa del dispositivo con el fin de documentarla. Se describe un caso raro de infección por Aspergillus fumigatus cinco años después de haber sido implantado un cardiodesfibrilador, se hace una revisión de la literatura y se repasan las principales características de la serie más amplia reportada.

Introducción: La estenosis aórtica severa impacta la morbilidad y mortalidad de los pacientes. Ésta se define como un área valvular menor a 1 cm2 o menor a 0,6 cm2/m2 de superficie corporal, gradientes transvalvulares mayores a 40 mm Hg y velocidades pico mayores a 4 m/s. Cuando los pacientes tienen estenosis aórtica severa y fracción de expulsión del ventrículo izquierdo conservada, se espera que estos valores sean elevados, pero se ha encontrado un subgrupo donde estos valores son menores, hecho que ha llevado a revaluar los puntos de corte para definir dicha enfermedad. Objetivo: Evaluar la prevalencia de estenosis aórtica severa con fracción de expulsión preservada de bajo flujo paradójico y determinar las características hemodinámicas e impacto clínico en los pacientes. Métodos: Estudio retrospectivo llevado a cabo durante 1999–2010, en un servicio de ecocardiografía, en el que se evaluaron 839 pacientes con diagnóstico de estenosis aórtica severa y fracción de eyección del ventrículo izquierdo conservada. La muestra fue subdividida en dos grupos: flujo normal definido como un volumen latido indexado (VLI) > 35 mL/m2 y grupo bajo flujo paradójico con un VLI ≤ 35 mL/m2 y se evaluaron sus características hemodinámicas, valvulares y de geometría ventricular, además de la supervivencia. Resultados: El 49,1% se clasificó como bajo flujo paradójico y el 50,9% como flujo normal. La edad promedio de fue 66,4 años y el 56,1% eran mujeres. La presión arterial media promedio fue 92,5 mm Hg, el gradiente medio 41,2 mm Hg y la fracción de expulsión promedio de 60%. Al comparar ambos grupos se encontró que los pacientes con bajo flujo paradójico tenían mayor impedancia válvulo-arterial (6,2 versus 4,25 mm Hg/mL/m, p

El síndrome de twiddler y su variante el síndrome de reel son complicaciones poco frecuentes en los pacientes portadores de dispositivos como marcapasos y cardiodesfibriladores implantables. Constituye una de las causas de disfunción del dispositivo, que puede conllevar complicaciones graves en quienes lo portan. Por tanto, se recomienda realizar evaluaciones periódicas con telemetría del dispositivo y radiografía de tórax, que aportan información valiosa en cuanto a prevención de complicaciones, como se expone en el caso.

La tormenta eléctrica postinfarto agudo de miocardio es una entidad amenazante de la vida y puede estar desencadenada por la isquemia activa secundaria a enfermedad no revascularizada. Adicionalmente la injuria miocárdica y sobre el sistema de conducción puede desencadenar la aparición de extrasístoles ventriculares, las cuales pueden participar como desencadenantes de arritmias ventriculares complejas. La adecuada identificación de las extrasístoles y otros posibles desencadenantes de la taquicardia ventricular son importantes para establecer las acciones terapéuticas que lleven a un adecuado control de la tormenta. Se presenta un caso de un paciente con tormenta arrítmica desencadenada por extrasístole ventricular frecuente con acoplamiento corto, la cual presentaba fenómeno de R en T, y era refractaria a manejo farmacológico, por lo que fue tratada exitosamente con ablación con catéter de radiofrecuencia.

la proteína C reactiva (PCR) es un predictor de eventos cardiovasculares, cuya relación con la muerte, el reinfarto o la revascularización luego del implante de stent en pacientes con síndrome coronario agudo, es un tema de investigación actual. Objetivo: evaluar el valor pronóstico de la proteína C reactiva (PCR) en pacientes con síndrome coronario agudo que se llevaron a intervencionismo percutáneo. Metodología: estudio retrospectivo en el que se comparó muerte, reinfarto o revascularización en 1, 6 y 12 meses después del intervencionismo percutáneo en el grupo de PCR baja (menor a 5 mg/ L) vs. PCR alta (mayor o igual a 5). Se aplicaron pruebas paramétricas para la comparación de dos medias (T de student) y Chi-cuadrado de Pearson. Resultados: se incluyeron 103 pacientes con síndrome coronario agudo, 60,2% con infarto sin elevación del ST y 41 con infarto con elevación del ST; la edad promedio fue 65, 4 ± 12,4 años y el valor promedio de la PCR de 7,7 ± 8,7 mg/L. No se encontraron diferencias respecto al MACE 1, 6 y 12 meses; se observó una tendencia no significativa a mayor muerte e infarto a seis meses y a revascularización e infarto a los doce meses en el grupo de PCR alta. Conclusión: los resultados sugieren una tendencia no significativa al aumento de los desenlaces cardiovasculares adversos en los pacientes con implante de stent por síndrome coronario agudo que tenían PCR alta.

BACKGROUND: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. METHODOLOGY/FINDINGS: An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05-0.5 parasites/mL whereas specific kDNA tests detected 5.10(-3) par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/µl of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/µl for each DNA stock, 0.5 par/mL and a sensitivity between 83.3-94.4%, specificity of 85-95%, accuracy of 86.8-89.5% and kappa index of 0.7-0.8 compared to consensus PCR reports of the 16 good performing tests and 63-69%, 100%, 71.4-76.2% and 0.4-0.5, respectively compared to serodiagnosis. Method LbD2 used solvent extraction followed by Sybr-Green based Real time PCR targeted to Sat-DNA; method LbD3 used solvent DNA extraction followed by conventional PCR targeted to Sat-DNA. The third method (LbF1) used glass fiber column based DNA extraction followed by TaqMan Real Time PCR targeted to Sat-DNA (cruzi 1/cruzi 2 and cruzi 3 TaqMan probe) and the fourth method (LbQ) used solvent DNA extraction followed by conventional hot-start PCR targeted to kDNA (primer pairs 121/122). These four methods were further evaluated at the coordinating laboratory in a subset of human blood samples, confirming the performance obtained by the participating laboratories. CONCLUSION/SIGNIFICANCE: This study represents a first crucial step towards international validation of PCR procedures for detection of T. cruzi in human blood samples.