Direct agglutination test for Encephalitozoon cuniculi

Serological Survey for Antibodies to Encephalitozoon cuniculi in Ownerless Dogs From Urban Areas of Brazil and Colombia

“…The sensitivity of the DAG in mice experimentally infected with the same strain of E. cuniculi used in the present study was 86% and the specificity was 98% (Jordan et al, 2006). These values are higher than the 68% sensitivity and 89% specificity observed in sera from naturally exposed dogs in the DAG in the present study.…”

“…Encephalitozoon cuniculi (ATCC 50502 ''canine subtype,'' Manassas, Virginia) was grown in human foreskin fibroblasts (Hs68, ATCC CRL1635, Manassas, Virginia) and the spores were harvested from the supernatant. Dog samples from Colombia were screened by the direct agglutination test (DAG) (Jordan et al, 2006) at a dilution of 1:50 in phosphate-buffered saline (PBS) using positive and negative control samples from a previous experiment as controls for the DAG. When a positive dog was identified, it was re-examined by the immunofluorescent assay test (IFAT) at a concentration of 1:50.…”

“…The DAG test was conducted as previously described (Jordan et al, 2006). Briefly, spores of E. cuniculi were collected and fixed in 2 ml of 37% formaldehyde solution for 10-15 sec in a 15-ml conical centrifuge tube and then diluted with PBS (pH 7.4) up to 15 ml and stored at 4 C. The antigen solution was prepared by washing the fixed spores twice in PBS and then resuspending them in alkaline buffer-eosin solution (7.02 g NaCl; 3.09 g H 3 BO 3 ; 24 ml 1 N NaOH; 4 g horse serum albumin factor V; 50 mg eosin Y; 1.0 g sodium azide; distilled H 2 O to make 1 L; pH 8.7).…”

Importance of Nonenteric Protozoan Infections in Immunocompromised People

“…1) performed on fixed tissue sections may be useful (55). A direct agglutination test (DAT) is also available, which can detect anti-E. cuniculi antibodies (302).…”

Molecular Diagnostic Tests for Microsporidia

“…Serologic tests such as the enzyme-linked immunosorbent assay (ELISA), immunoblot, and agglutination-based tests [66, 67] which can detect circulating antibody are not currently recommended for diagnostic purposes due to variable expression of antibodies in immunocompromised patients, the inability to discriminate between acute and past infections [68], the high prevalence of anti-microsporidian antibodies in apparently healthy, immunocompetent populations [6, 7], and cross-reactivity of antibodies between different species. However, these serologic analyses may be useful to diagnose subclinical infections in prospective transplant donors or patients who may be at risk for reactivation of infection due to impending immune compromise (discussed in [8]).…”