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Coccidioides species determination: does sequence analysis agree with restriction fragment length polymorphism?

Suzanne M Johnson¹, Erin L Carlson, Demosthenes Pappagianis

¹Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis, CA, 95616, USA, smjohnson@ucdavis.edu.

|January 12, 2015

Summary

Genetic analysis of Coccidioides isolates using restriction fragment length polymorphism (RFLP) and gene sequencing confirmed species division. Most RFLP groups corresponded to Coccidioides posadasii and Coccidioides immitis, with one potential hybrid identified.

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Area of Science:

Mycology
Molecular Biology
Genetics

Background:

Coccidioides fungal isolates exhibit genetic diversity.
Previous restriction fragment length polymorphism (RFLP) analysis divided 15 isolates into two groups.
Recent molecular studies established two distinct Coccidioides species: Coccidioides posadasii and Coccidioides immitis.

Purpose of the Study:

To test the hypothesis that RFLP groupings correlate with the established species division of Coccidioides.
To determine the species assignment for Coccidioides isolates using molecular methods.

Main Methods:

Amplification and sequencing of the ribosomal DNA internal transcribed spacer region and three genes (dioxygenase, serine proteinase, urease) from 13 Coccidioides isolates.
Phylogenetic analysis of sequencing data to assign species.
Comparison of species assignments with previous RFLP groupings.

Main Results:

Species assignment based on gene sequencing largely agreed with RFLP groupings.
One isolate showed discordance, suggesting it may be a hybrid.
Polymorphic sites across the four examined genes were consistent for species assignment.

Conclusions:

The study supports the correlation between RFLP groups and Coccidioides species (C. posadasii and C. immitis).
A potential hybrid Coccidioides isolate was identified.
Analysis of a single gene may be sufficient for accurate species identification within the Coccidioides genus.