Identification of Coccidioidomycosis Causative Agent
by Means of Polymerase Chain Reaction

G. A. Tkachenko, M. A. Grishina, V. A. Antonov, S. S. Savchenko,
V. S. Zamaraev, V. S. Lesovoi, and A. V. Lipnitskii

Volgograd Research Antiplague Institute, Volgograd, Russia

Received August 28, 2006

Abstract—For the purpose of diagnosing coccidioidomycosis by means of PCR, two pairs of primers were
constructed. The first one was developed on the basis of MBP-I gene to be used for identification of two Coc-
cidioides species, C. immitis and C. posadasii. The other pair of primers was constructed on the basis of the
gene encoding immunodominant glycoprotein of the spherule outer wall (SOWgp82) to be used for identifica-
tion of C. posadasii. With these primers, both highly sensitive and highly specific identification of coccidioido-
mycosis causative agents proved to be possible. An efficient procedure was developed for isolation of fungal
DNA from soil contaminated with Coccidioides spp. arthrospores. This procedure includes guanidine thiocy-
anate–phenol–chloroform deproteinization followed by DNA purification by means of nucleosorption.

DOI:10.3103/S0891416807040052

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