ELISA method for detecting microsporidia.

Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.

ELISA method for detecting microsporidia.

Natl Conf Hum Retroviruses Relat Infect (1st). 1993 Dec 12-16;:140. Unique Identifier : AIDSLINE AIDS/95921489
Didier ES; Aldras AM; Bertucci DC; Tulane Regional Primate Research Center, Covington, LA.

Abstract: Microsporidia are increasingly recognized as causing opportunistic infections in AIDS patients. Microsporidia are difficult to diagnose microscopically because the organisms are quite small. The purpose of this study was to develop an ELISA for detecting microsporidia in stool or urine specimens. To enrich for microsporidia, the formalin-fixed stool specimens were centrifuged in 50% Percoll, and resuspended in the original volume with tris-buffered saline (TBS). Urine sediment was obtained by centrifugation and resuspension in 1/100th of the original volume in TBS. The most consistent results were obtained when the specimens were allowed to dry onto ELISA plates (50 microliter per well), followed by fixation with a mixture of equal volumes of acetone and methanol. The ELISA plates were then sequentially incubated with hyperimmune rabbit polyclonal antisera raised against the microsporidia, goat anti-rabbit IgG, alkaline phosphatase-conjugated rabbit anti-goat, and p-nitrophenyl phosphate with washing in between each step. To reduce non-specific background, we found it necessary to absorb the primary antiserum with a control stool specimen spiked with Candida albicans. In addition, it was necessary to absorb the secondary and tertiary antibodies with E. cuniculi for comparing results with negative control serum. Using this method, we were able to detect as few as 1000 formalin-fixed tissue culture-derived Encephalitozoon hellem as well as Enterocytozoon bieneusi in stool diluted as far as 1:300. Further studies are required to fully assess sensitivity, specificity and reliability of this ELISA method. However, this ELISA should prove useful in screening large numbers of stool and urine specimens for detecting microsporidia infections.

Keywords: Fluorescent Antibody Technique Human HIV Seropositivity/*COMPLICATIONS Prognosis Retrospective Studies Serodiagnosis Toxoplasmosis, Cerebral/*DIAGNOSIS ABSTRACT
951230
M95C1660

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