Orally transmitted Chagas disease (ChD), which is a well-known entity in the Brazilian Amazon Region, was first documented in Venezuela in December 2007, when In Venezuela, effective control measures successfully reduced the annual incidence of Trypanosoma cruzi infections from 10/1,000 individuals in the 1950s to 1/1,000 in the 1980s (Feliciangeli et al. 2003). Since then, the recorded outbreaks of acute Chagas disease (ChD) have been as follows: between 1988-1996, 59 cases occurred in the western states of Venezuela (Añez et al. 1999), after which nine cases involving two deaths occurred during the period of January 2006-March 2007 in the same endemic area (Añez et al. 2007). In 2004 and 2006, the deaths of two children near Caracas were documented ). Nevertheless, an outbreak of orally transmitted ChD (OChD) in a school in a middle class neighbourhood in Caracas was the warning signal that ChD was re-emerging via a different mode of transmission, i.e., the oral route (Alarcón de Noya et al. 2010b Noya et al. 2010b), an outbreak occurred in an urban school that was presumably linked to the ingestion of contaminated home-made juice that had been prepared in a different part of the city where triatomines were present.The aim of this study is to provide information about the parasitological, immunological and molecular laboratory tests used in diagnosing ChD to gleam further knowledge from the largest OChD outbreak ever reported.
SUBJECTS, MATERIALS AND METHODSStudy population -The study population consisted of students, teachers and administrative personnel from a school located in the municipality of Chacao in Caracas, Venezuela. Once the primary case was diagnosed (Martín et al. 2009) and the epidemiological link to another case was established (Alarcón de Noya et al. 2010c), parasitological examinations and serological tests were carried out in these cases, in addition to other symptomatic patients who voluntarily requested medical attention at the Tropical Medicine Institute (IMT), Universidad Central de Venezuela in Caracas. After the death of a five-year-old student, most of the school community was managed as part of an on-site emergency, both in the school and at the hospital, where several of the patients were hospitalised. T. cruzi infection was diagnosed by collecting only one tube of blood for serological testing, except for in people who were symptomatic at the time of the interview and from whom additional blood was taken for parasitological or molecular testing (Supplementary data). For logistical and ethical reasons, a unique protocol could not be followed and, consequently, the results are simply presented as they were obtained over time. It was not possible to re-sample the blood of individuals with positive serological tests to isolate the parasite before treatment because the treatment had to begin before the group dispersed for the Christmas holiday.Direct parasitological tests -Observations of the parasite were performed using fresh blood smears as well as Giemsa-stained preparations (W...