Typing of Histoplasma capsulatum by restriction fragment length polymorphisms in a nuclear gene.
We previously described yps-3, a Histoplasma-specific nuclear gene probe useful in the identification of Histoplasma capsulatum. By using restriction fragment length polymorphisms (RFLPs) of DNA detected by the yps-3 gene and mitochondrial DNA, 76 clinical and soil isolates of H. capsulatum were classified. The majority of North American isolates obtained from endemic regions of the midwestern United States were members of the previously characterized class 2, although four clinical isolates from different patients with AIDS from that region were grouped in class 1 with the temperature-sensitive Downs strain. A Florida soil isolate (FLS1) was placed in class 4 on the basis of RFLP with both probes. Two American Type Culture Collection strains (G184B and G186B) from Panama were grouped into class 3 by this analysis. A group of five H. capsulatum isolates obtained from patients with AIDS in New York City were typed into a new class 5 on the basis of yps-3 polymorphisms; those organisms fell into two broad mitochondrial DNA patterns, designated 5b and 5c. Two new isolates from Panama were also members of this broad yps-3 class 5 group, but they exhibited a distinct mitochondrial DNA profile (class 5a). A sixth class was detected in DNA obtained from a patient with AIDS from Panama; that DNA had unique RFLP profiles with respect to both probes. These observations suggest that the Histoplasma-specific yps-3 gene probe is a sensitive tool for typing H. capsulatum in clinical specimens. Additionally, these studies provide molecular support for the hypothesis that AIDS-associated histoplasmosis in nonendemic areas is due to reactivation of a previously acquired infection.
Keath
E J
EJ
Department of Biology, Saint Louis University, Missouri 63103.
Kobayashi
G S
GS
Medoff
G
G
D000163:Acquired Immunodeficiency Syndrome / Q000150:complications; D015342:DNA Probes; D004271:DNA, Fungal / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D005069:Evaluation Studies as Topic; D005800:Genes, Fungal; D006658:Histoplasma / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006660:Histoplasmosis / Q000150:complications / Q000382:microbiology; D006801:Humans; D009172:Mycology / Q000379:methods; D009894:Opportunistic Infections / Q000150:complications / Q000382:microbiology; D012150:Polymorphism, Restriction Fragment Length
D015342:DNA Probes; D004271:DNA, Fungal; D004272:DNA, Mitochondrial