Evaluation of an immunodot blot technique for the detection of antibodies against Taenia solium larval antigens.

Immunodiagnostic tests represent an important tool for diagnosis of cysticercosis, the disease caused by cysticerci of Taenia solium. Accurate diagnosis of neurocysticercosis (NCC) requires costly neuroimaging techniques (magnetic resonance imaging and computed tomography), which are seldom affordable for people in endemic countries. Hence, new low-cost diagnostic methods offering good sensitivity and specificity are needed. Here, we studied four immunodiagnostic tests immunodot blot Tsol-p27, a commercial ELISA, and Western blot Tsol-p27/TsolHSP36, and compared them with a commercial enzyme-linked immunoelectrotransfer blot (EITB) that we regarded as the gold standard method. The analyzed serum samples were obtained from 160 patients: 94 epileptics suspected of NCC, six individuals confirmed NCC-positive, and 60 with positive (30) or negative (30) serology for Chagas diseases. Of the 100 serum samples from epileptic patients, 13 were positive and 87 negative by EITB. Compared to Western blot Tsol-p27, immunodot blot Tsol-p27 offered similar specificity (97.8% vs. 95.6%) but better sensitivity (86.7% vs. 76.4%). The ELISA was similar to the immunodot blot Tsol-p27 regarding both sensitivity and specificity. Western blot TsolHSP36 provided the lowest sensitivity (61.9%) and specificity (86.1%). None of the antibodies in the serum samples from the Chagas control groups were recognized by immunodot blot Tsol-p27. Our results indicate that the immunodot blot Tsol-p27 provides good sensitivity and specificity. Furthermore, considering the simplicity and low cost of this test, it might be preferable as a diagnostic method in poorly equipped laboratories in endemic countries.

Dominance of Giardia assemblage B in León, Nicaragua.

Giardiasis is a major problem in León, Nicaragua, yet despite this no data are available regarding the prevalence of different Giardia genotypes in this area. To address this question, a molecular analysis of Giardia isolates from humans and dogs living in the same area in León, Nicaragua was performed. Giardia isolates from 119 Nicaraguan patients and 8 dogs were successfully genotyped using single and/or nested beta-giardin PCR with subsequent restriction length fragment polymorphism (RFLP) analysis. The analyses of human samples yielded 94 (79%) assemblage B isolates and 25 (21%) assemblage A isolates. Only the non-human-associated assemblages C and D were found in the dog samples. Sixteen isolates with assemblage A pattern, 26 isolates with assemblage B pattern and all dog isolates were further characterized by sequencing the nested beta-giardin PCR product and by molecular analyses of the glutamate dehydrogenase (gdh) gene. Within the study area the assemblage A isolates were highly genetically homogenous, showing only sub-genotypes A2 (n=3) or A3 (n=13) at the beta-giardin locus and AII only at the gdh locus while assemblage B showed a high genetic polymorphism at both loci. Seven different sub-genotypes were identified within 13 of the sequenced assemblage B beta-giardin isolates. The remaining 13 sequenced assemblage B-isolates appeared to contain several different variants of the beta-giardin gene since the chromatograms displayed one to seven double peaks. The gdh sequences showed an even higher polymorphism since only 2 of 26 assemblage B isolates were without double peaks. Two mixed infections between assemblage A and B were found when the gdh gene was analyzed. Polymorphisms were also observed in the dog-associated assemblages C and D, but to a lesser extent than in assemblage B.

Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study.

BACKGROUND: Based on stool microscopy, an E. histolytica/E. dispar prevalence of 18.6% was found in León, Nicaragua about 10 years ago. Since then, new non-microscopic methods have been developed to discriminate between pathogenic E. histolytica and nonpathogenic E. dispar. The main objectives of the present study were to evaluate the true prevalence of E. histolytica among individuals with diarrhea and to assess the diagnostic procedures carried out at the health center level. METHODS: A descriptive study was carried out on patients with diarrhea. Parasite detection was performed by conventional microscopy on native preparations or concentrated and stained specimens, Triage Parasite Panel and by PCR for both E. histolytica and E. dispar. RESULTS: In 134 individuals with diarrhea, the prevalence of intestinal parasites was 69% as detected by direct stool examination. E. histolytica/E. dispar was found in eight (6%) of the samples, but the health centers reported 24%. In the Triage Parasite Panel only one case of E. histolytica/E. dispar was found. Analysis by PCR showed E. dispar in ten (7.5%) and E. histolytica in two cases (1.5%). The detection of intestinal coccidia and Dientamoeba fragilis required additional staining methods. CONCLUSIONS: PCR results showed that E. histolytica is a rare finding in patients with diarrhea. At the health centers, E. histolytica, E. histolytica/E. dispar were clearly overdiagnosed, with the consequence of overtreatment.

Antibodies in mother's milk protect children against giardiasis.

The protective effect of anti-Giardia antibodies in mother's milk on the acquisition of Giardia infection in their children during the first 2 y of life was analysed as part of a prospective study on infant diarrhoea in a group of 307 mothers and children in Leòn, Nicaragua. Among 24 children acquiring infection within the first 6 months, 23 were born to mothers lacking antibodies. These children also developed more severe diarrhoea. A significant difference between children born to mothers with and without antibodies with respect to the age at which the first Giardia infection was acquired was demonstrated by survival analysis and log rank test (p = 0.036). In conclusion, children born to non-immune mothers are at significantly higher risk of acquiring Giardia infection and developing giardiasis with more severe symptoms compared with children of immune mothers.