Dataset Information

Design and evaluation of an AFLP molecular marker for the detection of Coccidioides spp. in biological samples.

ABSTRACT: At present, there is no standardized marker that is routinely used in clinical laboratories to diagnose coccidioidomycosis. Thus, the goals of this study were to obtain a sequence characterized amplified region (SCAR) marker for the identification of Coccidioides spp., evaluate its specificity and sensitivity in fungal DNA-spiked blood and sputum samples, and compare it with previously described molecular markers. Specific amplified fragment length polymorphism (AFLP) amplicons for Coccidioides immitis and Coccidioides posadasii were cloned into the vector pGEM® -T Easy vector and sequenced to develop a SCAR marker. Oligonucleotides were designed to identify Coccidioides spp. by polymerase chain reaction (PCR), and the specificity and sensitivity of these oligonucleotides were tested with the DNA from related pathogens. The specificity and sensitivity of the SCAR marker was evaluated with blood and sputum samples spiked with Coccidioides DNA and compared with other previously described markers (621, GAC2, and Ag2/PRA). In addition, the conditions for its use were established using biological samples. A specific marker named SCAR₃₀₀ was obtained to identify Coccidioides spp. that exhibited good sensitivity and specificity. The results showed that all of the markers tested in this study can identify Coccidioides spp. However, the SCAR₃₀₀ and 621 markers were the most sensitive, whereas the SCAR₃₀₀ marker was the most specific. Thus, the SCAR₃₀₀ marker is a useful tool to identify Coccidioides spp.

SUBMITTER: Reyes-Montes MDR

PROVIDER: S-EPMC9428003 | biostudies-literature | 2019 Sep - Oct

REPOSITORIES: biostudies-literature

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Design and evaluation of an AFLP molecular marker for the detection of Coccidioides spp. in biological samples.

Reyes-Montes María Del Rocío MDR Frías-De-León María Guadalupe MG Victoriano-Pastelín Isai I Acosta-Altamirano Gustavo G Duarte-Escalante Esperanza E

The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases 20190917 5

At present, there is no standardized marker that is routinely used in clinical laboratories to diagnose coccidioidomycosis. Thus, the goals of this study were to obtain a sequence characterized amplified region (SCAR) marker for the identification of Coccidioides spp., evaluate its specificity and sensitivity in fungal DNA-spiked blood and sputum samples, and compare it with previously described molecular markers. Specific amplified fragment length polymorphism (AFLP) amplicons for Coccidioides ...[more]

PMID: 31539511

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Dataset Information

Design and evaluation of an AFLP molecular marker for the detection of Coccidioides spp. in biological samples.

Publications

Design and evaluation of an AFLP molecular marker for the detection of Coccidioides spp. in biological samples.

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