Zaida E. Yadon

Chagas disease: a Latin American health problem becoming a world health problem.

Political repression and/or economic stagnation stimulated the flow of migration from the 17 Latin American countries endemic for Chagas disease to developed countries. Because of this migration, Chagas disease, an autochthonous disease of the Continental Western Hemisphere is becoming a global health problem. In 2006, 3.8% of the 80,522 immigrants from those 17 countries to Australia were likely infected with Trypanosoma cruzi. In Canada in 2006, 3.5% of the 156,960 immigrants from Latin America whose country of origin was identified were estimated to have been infected. In Japan in 2007, there were 80,912 immigrants from Brazil, 15,281 from Peru, and 19,413 from other South American countries whose country of origin was not identified, a portion of whom may have been also infected. In 15 countries of Europe in 2005, excluding Spain, 2.9% of the 483,074 legal Latin American immigrants were estimated to be infected with T. cruzi. By 2008, Spain had received 1,678,711 immigrants from Latin American endemic countries; of these, 5.2% were potentially infected with T. cruzi and 17,390 may develop Chagas disease. Further, it was estimated that 24-92 newborns delivered by South American T. cruzi infected mothers in Spain may have been congenitally infected with T. cruzi in 2007. In the USA we estimated that 1.9% of approximately 13 million Latin American immigrants in 2000, and 2% of 17 million in 2007, were potentially infected with T. cruzi. Of these, 49,157 and 65,133 in 2000 and 2007 respectively, may have or may develop symptoms and signs of chronic Chagas disease. Governments should implement policies to prevent donations of blood and organs from T. cruzi infected donors. In addition, an infrastructure that assures detection and treatment of acute and chronic cases as well as congenital infection should be developed.

Analytical Performance of a Multiplex Real-Time PCR Assay Using TaqMan Probes for Quantification of Trypanosoma cruzi Satellite DNA in Blood Samples

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, 4- 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 Chagas disease patients under etiological treatment.

Elimination of Neglected Diseases in Latin America and the Caribbean: A Mapping of Selected Diseases

In Latin America and the Caribbean, around 195 million people live in poverty, a situation that increases the burden of some infectious diseases. Neglected diseases, in particular, are often restricted to poor, marginalized sections of the population. Tools exist to combat these diseases, making it imperative to work towards their elimination. In 2009, the Pan American Health Organization (PAHO) received a mandate to support the countries in the Region in eliminating neglected diseases and other poverty-related infections. The objective of this study is to analyze the presence of selected diseases using geo-processing techniques. Five diseases with information available at the first sub-national level (states) were mapped, showing the presence of the disease (“hotspots”) and overlap of diseases (“major hotspots”). In the 45 countries/territories (approximately 570 states) of the Region, there is: lymphatic filariasis in four countries (29 states), onchocerciasis in six countries (25 states), schistosomiasis in four countries (39 states), trachoma in three countries (29 states), and human rabies transmitted by dogs in ten countries (20 states). Of the 108 states with one or more of the selected diseases, 36 states present the diseases in overlapping areas (“major hotspots”). Additional information about soil-transmitted helminths was included. The analysis suggests a majority of the selected diseases are not widespread and can be considered part of an unfinished agenda with elimination as a goal. Integrated plans and a comprehensive approach, ensuring access to existing diagnostic and treatment methods, and establishing a multi-sectoral agenda that addresses social determinants, including access to adequate water and sanitation, are required. Future studies can include additional diseases, socio-economic and environmental variables.

Congenital Chagas disease: estimating the potential risk in the United States.

Economic hardship and/or political turmoil stimulated migration of Trypanosoma cruzi-infected population from Latin American countries to the United States and Europe; originating cases of Chagas disease were transmitted through blood, organ donation, and vertical transmission. Hispanic immigrant women of reproductive age in the United States coming from Chagas disease-endemic countries accounted for 2,384,644, and 5,841,538 in 1990 and 2000, respectively. Considering the prevalence rates for T. cruzi infection in their country of origin and the risk of newborns from infected mothers to acquire congenital infection as 1.33% and 5%, we estimated that the number of T. cruzi-infected newborns was 85-318 in 1990 and 166-638 in 2000. Diagnosis of infection in the mother and newborns at risk is needed. A high rate of cure is achieved, almost 100%, when the offspring is treated early. Health authorities, professional associations, physicians, and Hispanic groups should pay more attention to the subject.

Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients

An international study was performed by 26 experienced PCR laboratories from 14 countries to assess the performance of duplex quantitative real-time PCR (qPCR) strategies on the basis of TaqMan probes for detection and quantification of parasitic loads in peripheral blood samples from Chagas disease patients. Two methods were studied: Satellite DNA (SatDNA) qPCR and kinetoplastid DNA (kDNA) qPCR. Both methods included an internal amplification control. Reportable range, analytical sensitivity, limits of detection and quantification, and precision were estimated according to international guidelines. In addition, inclusivity and exclusivity were estimated with DNA from stocks representing the different Trypanosoma cruzi discrete typing units and Trypanosoma rangeli and Leishmania spp. Both methods were challenged against 156 blood samples provided by the participant laboratories, including samples from acute and chronic patients with varied clinical findings, infected by oral route or vectorial transmission. kDNA qPCR showed better analytical sensitivity than SatDNA qPCR with limits of detection of 0.23 and 0.70 parasite equivalents/mL, respectively. Analyses of clinical samples revealed a high concordance in terms of sensitivity and parasitic loads determined by both SatDNA and kDNA qPCRs. This effort is a major step toward international validation of qPCR methods for the quantification of T. cruzi DNA in human blood samples, aiming to provide an accurate surrogate biomarker for diagnosis and treatment monitoring for patients with Chagas disease.

Interventions for American Cutaneous and Mucocutaneous Leishmaniasis: A Systematic Review Update

Introduction Leishmaniasis is an important public health problem in the Americas. A Cochrane review published in 2009 analyzed 38 randomized controlled trials (RCT). We conducted a systematic review to evaluate the effects of therapeutic interventions for American cutaneous and mucocutaneous leishmaniasis. Methods All studies were extracted from PubMed, Embase, Lilacs (2009 to July, 2012 respectively), the Cochrane Central Register of Controlled Trials (6-2012) and references of identified publications. RCTs’ risk of bias was assessed. Results We identified 1865 references of interest; we finally included 10 new RCTs. The risk of bias scored low or unclear for most domains. Miltefosine was not significantly different from meglumine antimoniate in the complete cure rate at 6 months (4 RCT; 584 participants; ITT; RR: 1.12; 95%CI: 0.85 to 1.47; I2 78%). However a significant difference in the rate of complete cure favoring miltefosine at 6 months was found in L. panamensis and L. guyanensis (2 RCTs, 206 participants; ITT; RR: 1.22; 95%CI: 1.02 to 1.46; I2 0%). One RCT found that meglumine antimoniate was superior to pentamidine in the rate of complete cure for L. braziliensis (80 participants, ITT; RR: 2.21; 95%CI: 1.41 to 3.49), while another RCT assessing L. guyanensis did not find any significant difference. Although meta-analysis of three studies found a significant difference in the rate of complete cure at 3 months favoring imiquimod versus placebo (134 participants; ITT; RR: 1.45; 95%CI: 1.12 to 1.88; I2 0%), no significant differences were found at 6 and 12 months. Thermotherapy and nitric oxide were not superior to meglumine antimoniate. Conclusion Therapeutic interventions for American cutaneous and mucocutaneous leishmaniasis are varied and should be decided according to the context. Since mucosal disease is the more neglected form of leishmaniasis a multicentric trial should be urgently considered.

Multiplex Real-Time PCR Assay Using TaqMan Probes for the Identification of Trypanosoma cruzi DTUs in Biological and Clinical Samples

Background Trypanosoma cruzi has been classified into six Discrete Typing Units (DTUs), designated as TcI–TcVI. In order to effectively use this standardized nomenclature, a reproducible genotyping strategy is imperative. Several typing schemes have been developed with variable levels of complexity, selectivity and analytical sensitivity. Most of them can be only applied to cultured stocks. In this context, we aimed to develop a multiplex Real-Time PCR method to identify the six T. cruzi DTUs using TaqMan probes (MTq-PCR). Methods/Principal Findings The MTq-PCR has been evaluated in 39 cultured stocks and 307 biological samples from vectors, reservoirs and patients from different geographical regions and transmission cycles in comparison with a multi-locus conventional PCR algorithm. The MTq-PCR was inclusive for laboratory stocks and natural isolates and sensitive for direct typing of different biological samples from vectors, reservoirs and patients with acute, congenital infection or Chagas reactivation. The first round SL-IR MTq-PCR detected 1 fg DNA/reaction tube of TcI, TcII and TcIII and 1 pg DNA/reaction tube of TcIV, TcV and TcVI reference strains. The MTq-PCR was able to characterize DTUs in 83% of triatomine and 96% of reservoir samples that had been typed by conventional PCR methods. Regarding clinical samples, 100% of those derived from acute infected patients, 62.5% from congenitally infected children and 50% from patients with clinical reactivation could be genotyped. Sensitivity for direct typing of blood samples from chronic Chagas disease patients (32.8% from asymptomatic and 22.2% from symptomatic patients) and mixed infections was lower than that of the conventional PCR algorithm. Conclusions/Significance Typing is resolved after a single or a second round of Real-Time PCR, depending on the DTU. This format reduces carryover contamination and is amenable to quantification, automation and kit production.

Eco-bio-social research on community-based approaches for Chagas disease vector control in Latin America

This article provides an overview of three research projects which designed and implemented innovative interventions for Chagas disease vector control in Bolivia, Guatemala and Mexico. The research initiative was based on sound principles of community-based ecosystem management (ecohealth), integrated vector management, and interdisciplinary analysis. The initial situational analysis achieved a better understanding of ecological, biological and social determinants of domestic infestation. The key factors identified included: housing quality; type of peridomestic habitats; presence and abundance of domestic dogs, chickens and synanthropic rodents; proximity to public lights; location in the periphery of the village. In Bolivia, plastering of mud walls with appropriate local materials and regular cleaning of beds and of clothes next to the walls, substantially decreased domestic infestation and abundance of the insect vector Triatoma infestans. The Guatemalan project revealed close links between house infestation by rodents and Triatoma dimidiata, and vector infection with Trypanosoma cruzi. A novel community-operated rodent control program significantly reduced rodent infestation and bug infection. In Mexico, large-scale implementation of window screens translated into promising reductions in domestic infestation. A multi-pronged approach including community mobilisation and empowerment, intersectoral cooperation and adhesion to integrated vector management principles may be the key to sustainable vector and disease control in the affected regions.

Target Product Profile (TPP) for Chagas Disease Point-of-Care Diagnosis and Assessment of Response to Treatment

A first and critical step to address the research and development gap for Chagas disease is to establish a consensus on the desirable product profiles in different conditions of use. To foster and inform the development of these much needed tools, the Pan American Health Organization (PAHO), in collaboration with DNDi, Medecins sans Frontieres (MSF), and the Special Programme for Research and Training in Tropical Diseases (TDR), convened a multidisciplinary group of experts in Rio de Janeiro, Brazil (April 2010), to review the evidence and initiate discussions. The meeting established the basis for the development of target product profiles (TPPs), which are reported in this paper.

Gains and Future Road Map for the Elimination of Dog-Transmitted Rabies in the Americas

In 1983 the countries of the Americas, with the technical cooperation of the Pan American Health Organization (PAHO), pledged to eliminate human rabies transmitted by dogs.1 Since then, countries have made great efforts to eliminate this disease, with notable success, within the framework of the Regional Program of Elimination of Human Rabies. The success achieved during the last 30 years and the historical solidarity between countries in the region support the goal of elimination of dog-transmitted rabies in the American continent by 2015. As this date approaches, there is a need to reflect and reassess the current plan of rabies elimination. This work briefly describes the achievements of the Regional Program and the proposed new Regional Action Plan for the elimination of dog-transmitted rabies.