Identification of novel diagnostic serum biomarkers for Chagas' disease in asymptomatic subjects by mass spectrometric profiling.

More than 10 million people are thought to be infected with Trypanosoma cruzi, primarily in the Americas. The clinical manifestations of Chagas' disease (CD) are variable, but most subjects remain asymptomatic for decades. Only 15 to 30% eventually develop terminal complications. All current diagnostic tests have limitations. New approaches are needed for blood bank screening as well as for improved diagnosis and prognosis. Sera from subjects with asymptomatic CD (n = 131) were compared to those from uninfected controls (n = 164) and subjects with other parasitic diseases (n = 140), using protein array mass spectrometry. To identify biomarkers associated with CD, sera were fractionated by anion-exchange chromatography and bound to two commercial ProteinChip array chemistries: WCX2 and IMAC3. Multiple candidate biomarkers were found in CD sera (3 to 75.4 kDa). Algorithms employing 3 to 5 of these biomarkers achieved up to 100% sensitivity and 98% specificity for CD. The biomarkers most useful for diagnosis were identified and validated. These included MIP1 alpha, C3a anaphylatoxin, and unusually truncated forms of fibronectin, apolipoprotein A1 (ApoA1), and C3. An antipeptide antiserum against the 28.9-kDa C terminus of the fibronectin fragment achieved good specificity (90%) for CD in a Western blot format. We identified full-length ApoA1 (28.1 kDa), the major structural and functional protein component of high-density lipoprotein (HDL), as an important negative biomarker for CD, and relatively little full-length ApoA1 was detected in CD sera. This work provides proof of principle that both platform-dependent (i.e., mass spectrometry-based) and platform-independent (i.e., Western blot) tests can be generated using high-throughput mass profiling.

Seroprevalence of Trypanosoma cruzi infection among school-age children in the endemic area of Guatemala.

In support of the National Program for Chagas Disease Control, we conducted a cross-sectional study to estimate the seroprevalence rate of Trypanosoma cruzi infection across the five Departments (Chiquimula, Jalapa, Zacapa, Jutiapa, and Santa Rosa) that are believed to comprise the entire principal endemic area in Guatemala. Also, so that the results could be used to identify areas of active transmission, we conducted the survey in school-aged children. We used an experimental enzyme-linked immunosorbent assay with blood samples obtained by finger prick to estimate the seroprevalence of T. cruzi. This assay has been previously tested and showed good sensitivity and specificity. Overall, the seropositivity rate for T. cruzi infection was 5.28% (235 of 4,450). Of 173 communities evaluated, 35 (20.23%) had a seropositive rate ranging from 10% to 45%. A number of parameters, including but not limited to living conditions, were examined for possible association with seropositivity. While there are several associations, the strongest association with seropositivity is living in a house with a thatch roof. The survey results will permit the Ministry of Health to stratify T. cruzi-endemic communities, enabling local health authorities to efficiently focus on vector control operations.

Molecular and serologic evidence of tick-borne Ehrlichiae in three species of lemurs from St. Catherines Island, Georgia, USA.

In recent years, several species of ehrlichiae have been recognized as tick-borne disease agents of veterinary and medical importance. Clinically normal free-ranging or previously free-ranging lemurs, including 46 ring-tailed lemurs (Lemur catta), six blue-eyed black lemurs (Eulemur macaco flavifrons), and four black and white ruffed lemurs (Varecia variegata variegata) from St. Catherines Island, Georgia, were tested for evidence of exposure to tick-borne ehrlichiae. All 52 adult lemurs were serologically tested for exposure to Ehrlichia chaffeensis and Anaplasma phagocytophilum. Polymerase chain reaction (PCR) assays for E. chaffeensis, A. phagocytophilum, Ehrlichia ewingii, and Ehrlichia canis were conducted on blood samples from all 56 lemurs. Blood from all lemurs was inoculated into DH82 cell cultures for E. chaffeensis isolation. Of the adult lemurs, 20 (38.5%) and 16 (30.8%) had antibodies reactive (> or =1:128) for E. chaffeensis and A. phagocytophilum, respectively. Two ring-tailed lemurs were PCR and culture positive for E. chaffeensis. Molecular characterization of the two E. chaffeensis isolates showed that both contained 5-repeat variants of the variable-length PCR target (VLPT) antigen gene and 3-repeat variants of the 120-kDa antigen gene. Sequencing of the VLPT genes revealed a novel amino acid repeat unit (type-9). One lemur infected with E. chaffeensis was slightly hypoproteinemic and had moderately elevated serum alanine aminotransferase (ALT) levels. These lemurs from St. Catherines Island have been exposed to or infected with tick-borne ehrlichiae, or both, but showed no clinical disease.

Prevalence of anti-Taenia solium antibodies in sera from outpatients in an Andean region of Ecuador

Sera from 9,254 individuals that presented at one of three outpatient clinics in Quito, Ecuador were assayed by indirect hemagglutination for the presence of antibodies reactive with antigens from Taenia solium cysts. Immunoblot anlysis of 81 selected sera with IHA titers ranging from 0 to 1,028 showed that a titer of maior ou igual a 32 was suggestive of exposure to the parasite. Nine percent (9 por cento) of the 9,254 patients had titers of 32 or greater. Of 3,503 sera from one clinic, which included sera from food handlers undergoing yearly physicals, 390 (11 por cento) were positive. In addition, a correlation with age was seen in some, but not all, populations. In situations where age-related effects were noted, the highest incidence was seen in the youngest (0-20 years) and in the oldest (51-60 years) group. Thus, a resurgence of infection after a period of lower prevalence may be developing. Overall, this study shows that cysticercosis is relatively common and potentially a serious health problem in this region.

Seroprevalence of Trypanosoma cruzi infection in three rural communities in Guatemala

A systematic, house-based serological survey for Trypanosoma cruzi seroreactivity was conducted in three contiguous communities in Olopa municipality, Chiquimula Department, Guatemala. Blood samples from a total of 292 individuals in 63 households were examined by enzyme-linked immunosorbent assay. The seropositive rate ranged from 0 x cent to 20.8 x cent for the three communities, with a mean of 15.1 x cent. Log-linear models showed that seroprevalence was significantly related to age (P0.005) but not to sex. However, when the age group with the lowest prevalence (1-9 years) was excluded from the analysis, age was not a significant factor (P=0.55). Data from a stratified sample collected at the same time were combined with those of the systematic sample to analyze the relationship between seropositivity and possible explanatory variables. Log-linear models, based on 586 individuals in 129 households from the two surveys, revealed a significant positive association between seropositivity and thatched roofs (P=0.01)

Seroprevalence of Trypanosoma cruzi infection in three rural communities in Guatemala / Seroprevalencia de infección por Trypanosoma cruzi en tres comunidades rurales de Guatemala

A systematic, house-based serological survey for Trypanosoma cruzi seroreactivity was conducted in three contiguous communities in Olopa municipality, Chiquimula Department, Guatemala. Blood samples from a total of 292 individuals in 63 households were examined by enzyme-linked immunosorbent assay. The seropositive rate ranged from 0 x cent to 20.8 x cent for the three communities, with a mean of 15.1 x cent. Log-linear models showed that seroprevalence was significantly related to age (P<0.005) but not to sex. However, when the age group with the lowest prevalence (1-9 years) was excluded from the analysis, age was not a significant factor (P=0.55). Data from a stratified sample collected at the same time were combined with those of the systematic sample to analyze the relationship between seropositivity and possible explanatory variables. Log-linear models, based on 586 individuals in 129 households from the two surveys, revealed a significant positive association between seropositivity and thatched roofs (P=0.01)

Una encuesta serológica sistemática y domiciliaria para detectar serorreactividad a Trypanosoma cruzi se llevó a cabo en tres comunidades contiguas de la municipalidad de Olapa, en el departamento de Chiquimula, Guatemala. Se examinaron mediante inmunoadsorción enzimática muestras de sangre obtenidas de un total de 292 habitantes que residían en 63 viviendas. La tasa de seropositividad varió de 0 a 20,8% en las tres comunidades y tuvo una media de 15,1%. Los modelos logarítmico-lineales mostraron una asociación significativa entre la seroprevalencia y la edad (P < 0,05), pero no entre aquella y el sexo. No obstante, cuando el grupo de edad que tuvo la seroprevalencia más baja (el de 1 a 9 años) se excluyó del análisis, la edad dejó de ser un factor significativo (P = 0,55). Datos obtenidos de una muestra estratificada recogida simultáneamente se combinaron con los de la muestra sistemática con el fin de analizar la relación entre la seropositividad y algunas variables que podrían ser explicativas. Los modelos logarítmico-lineales, aplicados en 586 habitantes de 129 viviendas incluidas en ambas encuestas, revelaron una asociación positiva significativa entre la seropositividad y la presencia de techo de paja (P = 0,01)