Speciation of human microsporidia by polymerase chain reaction single-strand conformation polymorphism.
We describe the application of single-strand conformation polymorphism (SSCP) analysis to the speciation of human microsporidia after polymerase chain reaction (PCR) amplification with the panmicrosporidian primers PMP1 and PMP2. We compared the DNA extracted and amplified from different genotypes or isolates of Enterocytozoon bieneusi, Encephalitozoon cuniculi, E. hellem, and E. intestinalis plus an isolate of Vittaforma corneae. The PCR-SSCP, when performed at 20 degrees C, generated 2 bands in distinctive, reproducible patterns in polyacrylamide gels for each species of microsporidia tested, regardless of genotype or isolate. We found PCR-SSCP to be an easy and reproducible method for speciation of human microsporidia when the primer pair PMP1 and PMP2 is used.
Fedorko
D P
DP
Microbiology Service and Chemistry Service, Department of Laboratory Medicine, Warren C. Magnuson Clinical Center, National Institutes of Health, Bethesda, Maryland 20892-1508, USA. dfedorko@nih.gov
Nelson
N A
NA
Didier
E S
ES
Bertucci
D
D
Delgado
R M
RM
Hruszkewycz
A M
AM
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005243:Feces / Q000469:parasitology; D005784:Gene Amplification; D005838:Genotype; D006801:Humans; D008565:Membrane Proteins; D016814:Microsporidia / Q000145:classification* / Q000235:genetics*; D016881:Microsporidiosis / Q000469:parasitology*; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D009419:Nerve Tissue Proteins; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011510:Proteolipids; D015203:Reproducibility of Results
D017931:DNA Primers; D016054:DNA, Protozoan; D008565:Membrane Proteins; D062806:Myelin and Lymphocyte-Associated Proteolipid Proteins; D009419:Nerve Tissue Proteins; C048897:PLLP protein, human; D011510:Proteolipids