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Detection and species identification of intestinal microsporidia in duodenal biopsies from HIV infected patients by polymerase chain reaction and Southern blot analysis.
David F; Schuitema AR; Sarfati C; Liguory O; Hartskeerl RA; Derouin F;
November 30, 1996
3rd Conf Retro and Opportun Infect. 1996 Jan 28-Feb 1;:160. Unique

Microsporidian infections are increasingly recognized among HIV infected patients. The standard method of characterization of the species involved,is transmission electron microscopy (TEM). Polymerase chain reaction (PCR) and Southern blot analysis (SB) was performed on duodenal biopsies obtained from 27 HIV infected patients in order to develop a sensitive and specific method for a routine diagnosis of microsporidian infections in HIV infected patients. Fifteen patients were infected with intestinal microsporidia. TEM identified E. bieneusi (EB) infection in 14 patients and E. intestinalis (EI) in one patient. Twelve patients not infected with microsporidia (negative stool and histological examination) served as controls, DNA prepared from EB and EI were used as PCR and SB positive controls. A primer set, amplifying EB, El, E.hellem and E. cuniculi was used for the PCR; two specific probes were used for species identification by SB, one for EB and one for EI. A 1300 bp DNA fragment could be amplified from 12 EB and from the EI infected biopsies (sensitivity = 87%); no DNA fragment could be amplified from biopsies without microsporidian infection (specificity = 100%). SB confirmed the TEM species identification from all positive PCR samples; in one sample SB showed a co-infection with EB and EI. These results suggest that PCR associated with SB is suitable for routine diagnosis and species identification of intestinal microsporidian infections in HIV infected patients.

Biopsy Blotting, Southern DNA, Protozoan Duodenum/PATHOLOGY/*PARASITOLOGY/ULTRASTRUCTURE Human Microscopy, Electron Microsporida/GENETICS/*ISOLATION & PURIF Polymerase Chain Reaction ABSTRACT

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