Cryptococcus spp. DNA extraction from environmental samples.
Keywords:
Cryptococcus spp., ecology, DNA extraction, PCR, environmental samples
Abstract
The genus Cryptococcus encompasses 38 species, but only 3 are associated with disease in humans and animals, Cryptococcus laurentii, Cryptococcus albidus and Cryptococcus neoformans. The last one is the most frequently reported. The disease is acquired by the inhalation of infectious propagules present in the environment. The habitat has been established using extraction techniques with buffer supplemented with antibiotics and plating in selective media. The aim of this work was to evaluate several DNA extraction techniques for Cryptocococus spp. from environmental samples. The control isolates were C. neoformans, C. albidus, C. laurentii and Paracoccidiodes brasiliensis. We also used vermiculita and soil samples contaminated with different yeast concentrations (10 to 10(6) cells/g) and samples naturally contaminated with C. neoformans. DNA was extracted with physical and chemical methods and with a commercial kit, and the DNA was purified with agarose blocks and silica columns. For the PCR amplification we used the CN4-CN5 primers, which are specific for C. neoformans. Only the commercial kit allowed DNA extraction and amplification from contaminated soil samples up to a concentration of 10 cells/g and from one sample naturally colonized. With this work we extracted and amplified DNA from Cryptococcus spp. from environmental samples with appropriate PCR specificity, it will be a tool to establish the ecological areas of C. neoformans in our country.Downloads
Download data is not yet available.
How to Cite
1.
Castañeda A, McEwen J, Hidalgo M, Castañeda E. Cryptococcus spp. DNA extraction from environmental samples. Biomed. [Internet]. 2004 Sep. 1 [cited 2024 Jul. 28];24(3):324-31. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/1279
Some similar items:
- Nohora M. Mendoza, Carlos A. Jaramillo, Felipe Guhl, Julio C. Padilla, Martha C. Rentería, Malaria diagnosis by nested PCR , Biomedica: Vol. 21 No. 4 (2001)
- Olga L. Cabrera, Leonard E. Munsterman, Rocío Cárdenas, Reynaldo Gutiérrez, Cristina Ferro, Definition of appropriate temperature and storage conditions in the detection of Leishmania DNA with PCR in phlebotomine flies. , Biomedica: Vol. 22 No. 3 (2002)
- Marylin Hidalgo, Claudia Parra, María Victoria Ovalle, Clara Inés Agudelo, Elizabeth Castañeda, Accuracy of the slide agglutination method evaluated with PCR in typing Haemophilus influenzae isolates. , Biomedica: Vol. 23 No. 2 (2003)
- Olga L. Cabrera, Leonard E. Munstermann, Rocío Cárdenas, Cristina Ferro, PCR as a tool in confirming the experimental transmission of Leishmania chagasi to hamsters by Lutzomyia longipalpis (Diptera:Psychodidae). , Biomedica: Vol. 23 No. 2 (2003)
- Miryan Margot Sánchez, Nora María Cardona-Castro, Development and evaluation of a PCR method for diagnosis of Salmonella enteric fever, based on DNA sequences of the hilA gene. , Biomedica: Vol. 24 No. 2 (2004)
- Jorge H. Botero, Martha Nelly Montoya, Adriana Lucía Vanegas, Abel Díaz, Luis Navarro-i-Martínez, Fernando Jorge Bornay, Fernando Izquierdo, Carmen del Aguila, Sonia del Pilar Agudelo, Frequency of intestinal microsporidian infections in HIV-positive patients, as diagnosis by quick hot Gram chromotrope staining and PCR. , Biomedica: Vol. 24 No. 4 (2004)
- Manzour Hernando Hazbón, Recent advances in molecular methods for early diagnosis of tuberculosis and drug-resistant tuberculosis. , Biomedica: Vol. 24 (2004): Suplemento 1
- Dora Fonseca, Andrés Gutiérrez, Claudia Silva, Mauricio Coll, Gustavo Malo, Camilo Orjuela, Alejandro Giraldo, Identification of point mutations in the 21-hydroxylase gene in patients affected with congenital adrenal hyperplasia. , Biomedica: Vol. 25 No. 2 (2005)
- Erika Santamaría, Nubia Ponce, Concepción Puerta, Cristina Ferro, Validation of PCR as a tool for the detection of Leishmania (Vianna) spp. parasites in the Lutzomyia (Diptera: Psychodidae) vector. , Biomedica: Vol. 25 No. 2 (2005)
- Angela Patricia Guerra, Angélica Knudson, Rubén Santiago Nicholls, John Alexander Galindo, Zaava Ravid, Sonia Rahirant, Nidia Duarte, Jacqueline Chaparro-Olaya, Moisés Wasserman, Genotyping of the Plasmodium falciparum msp1 (block 2) and dhfr (codon 108) genes in field samples collected in four endemic Colombian localities. , Biomedica: Vol. 26 No. 1 (2006)
Published
2004-09-01
Issue
Section
Technical note
Article metrics | |
---|---|
Abstract views | |
Galley vies | |
PDF Views | |
HTML views | |
Other views |