Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 May 16:11:108.
doi: 10.1186/1471-2180-11-108.

Molecular identification of Coccidioides spp. in soil samples from Brazil

Regina C L de Macêdo  1 , Alexandre S Rosado, Fabio F da Mota, Maria A S Cavalcante, Kelsen D Eulálio, Antônio D Filho, Liline M S Martins, Márcia S Lazéra, Bodo Wanke
Affiliations

Molecular identification of Coccidioides spp. in soil samples from Brazil

Regina C L de Macêdo et al. BMC Microbiol. .

Abstract

Background: Since 1991 several outbreaks of acute coccidioidomycosis (CM) were diagnosed in the semi-arid Northeast of Brazil, mainly related to disturbance of armadillo burrows caused by hunters while digging them for the capture of these animals. This activity causes dust contaminated with arthroconidia of Coccidioides posadasii, which, once inhaled, cause the mycosis. We report on the identification of C. posadasii in soil samples related to outbreaks of CM.

Results: Twenty four soil samples had their DNA extracted and subsequently submitted to a semi-nested PCR technique using specific primers. While only 6 (25%) soil samples were positive for C. posadasii by mice inoculation, all (100%) were positive by the molecular tool.

Conclusion: This methodology represents a simple, sensitive and specific molecular technique to determine the environmental distribution of Coccidioides spp. in endemic areas, but cannot distinguish the species. Moreover, it may be useful to identify culture isolates. Key-words: 1. Coccidioidomycosis. 2. Coccidioides spp. 3. C. posadasii. 4. Semi-arid. 5. Semi-nested PCR.

PubMed Disclaimer

Figures

Figure 1
Figure 1
1.2% agarose gel showing results of semi-nested PCR with primers RFA12/RFA13 and RFA12/P2 specifics for Coccidioides spp., lines 1-4 DNA isolated of C. immitis (US), lines 5-9 DNA isolated of C. posadasii (Piauí/Brazil), and line 10 negative control (DNA C. neoformans). MW = 1 Kb DNA Ladder (Promega).
Figure 2
Figure 2
1.2% agarose gel showing results of semi-nested PCR with primers RFA12/RFA13 and RFA12/P2 specifics for Coccidioides spp. lines 1-2 DNAs Rhodococcus equi 33701 e Mycobacterium avium 13956, lines 3-4 DNA isolated of C. immitis (US), lines 5-6 DNA isolated of C. posadasii (Argentina) and lines 7-13 DNA isolated of C. posadasii (Piauí/Brazil) MW = 1 Kb DNA Ladder (Promega).
Figure 3
Figure 3
1.2% agarose gel showing results of semi-nested PCR with primers RFA12/RFA13 and RFA12/P2 specific for Coccidioides spp., lines 2-11 soil samples from Elesbão Veloso (EV), lines 12 and 13 Caridade do Piauí (CP). Line 1 = white and MW = 1 Kb DNA Ladder (Promega).
Figure 4
Figure 4
1.2% agarose gel showing results of semi-nested PCR with primers RFA12/RFA13 and RFA12/P2 specific for Coccidioides spp., lines 1, 5, 9, 13 and 17 = white, lines 2-4 DNA C. posadasii (pure), lines 6-8 DNA C. posadasii (diluted at 10-2), lines 10-12 DNA C. posadasii (diluted a 10-3), lines 14-16 DNA C. posadasii (diluted 10-4). MW = 1 Kb DNA Ladder (Promega).
See this image and copyright information in PMC

References

    1. Fisher MC, Koenig GL, White TJ, Taylor JW. Molecular and phenotypic description of Coccidioides posadasii sp. nov., previously recognized as the non-California population of Coccidioides immitis. Mycologia. 2002;94(1):73–84. doi: 10.2307/3761847. - DOI - PubMed
    1. Pappagianis D. In: Current topics of medical mycology. McGinnis MR, editor. Vol. 2. Springer-Verlag, New York; 1988. Epidemiology of coccidioidomycosis; pp. 199–238. - PubMed
    1. Ajello L. Coccidioidomycosis and histoplasmosis: a review of its epidemiology and geographical distribution. Mycopathologia. 1971;45:221–230. - PubMed
    1. Hector RF, Laniado-Laborin R. Coccidioidomycosis -A fungal disease of the Americas. PloS Med. 2005;2(1):15–18. doi: 10.1371/journal.pmed.0020015. - DOI - PMC - PubMed
    1. Mayorga RP, Espinoza H. Coccidioidomycosis in México and Center America. Mycopath Mycol Appl. 1970. pp. 13–23. - PubMed

Publication types

LinkOut - more resources