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

Reyes-Montes, María del Roció; Frías-De-León, María Guadalupe; Victoriano-Pastelín, Isai; Acosta-Altamirano, Gustavo; Duarte-Escalante, Esperanza

doi:10.1016/j.bjid.2019.08.002

Cited by 2 publications

(1 citation statement)

References 31 publications

(42 reference statements)

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“…They were applied on various sample types, such as formalin‐fixed paraffin‐embedded and fresh biopsies, 4,32,40,41 blood, 28,42–45 serum, 29,33,43,45–47 bronchoalveolar lavage fluid (BAL), 28,45,46,48 cerebrospinal fluid, 28,45,47,49 bone marrow, 28,43,48,49 pleural fluid 28,48 and urine 50 . They target multicopy ribosomal DNAs 28,29,33,36,39,43–45,47,49,51–59 (rDNAs) or species‐specific genes, 27,41,42,46,48,60–73 mostly single copy, except for the region identified in the NCBI database as a ‘copia‐like retrotransposon’ 1,37,38,74,75 with approximately 60 copies per Coccidioides genome 74 …”

Section: Introductionmentioning

confidence: 99%

A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis

Wilmes

Hagen

Veríssimo

et al. 2023

Mycoses

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Background: In-house real-time PCR (qPCR) is increasingly used to diagnose the socalled endemic mycoses as commercial assays are not widely available.Objectives: To compare the performance of different molecular diagnostic assays for detecting Histoplasma capsulatum and Coccidioides spp. in five European reference laboratories.Methods: Two blinded external quality assessment (EQA) panels were sent to each laboratory that performed the analysis with their in-house assays. Both panels included a range of concentrations of H. capsulatum (n = 7) and Coccidioides spp. (n = 6), negative control and DNA from other fungi. Four laboratories used specific qPCRs, and one laboratory a broad-range fungal conventional PCR (cPCR) and a specific cPCR for H. capsulatum with subsequent sequencing.Results: qPCR assays were the most sensitive for the detection of H. capsulatum DNA.The lowest amount of H. capsulatum DNA detected was 1-4 fg, 0.1 pg and 10 pg for qPCRs, specific cPCR and broad-range cPCR, respectively. False positive results occurred with high concentrations of Blastomyces dermatitidis DNA in two laboratories and with Emergomyces spp. in one laboratory. For the Coccidioides panel, the lowest amount of DNA detected was 1-16 fg by qPCRs and 10 pg with the broad-range cPCR. One laboratory reported a false positive result by qPCR with high load of Uncinocarpus DNA.

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Section: Introductionmentioning

confidence: 99%

A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis

Wilmes

Hagen

Veríssimo

et al. 2023

Mycoses

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Clinical Cases of Coccidioidomycosis in the Americas in the Period 1950–2021: Epidemiology Data, Diagnosis, and Treatment

Reyes-Montes,

León-García,

Frías-De-León

et al. 2023

Life

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Coccidioidomycosis, caused by Coccidioides immitis and C. posadasii, causes significant morbidity and mortality, both in immunocompetent and immunocompromised people, mainly in endemic areas. The present work analyzed its epidemiology, diagnostic methods, and treatment by reviewing clinical cases published from 1950 to 2021. Fifty-nine articles were included, corresponding to 275 clinical cases. The results showed a higher incidence of coccidioidomycosis in the male gender than the female gender. The most affected age group was 31–40 years, and the most reported clinical presentation was disseminated with greater involvement in cutaneous and subcutaneous tissue, followed by the CNS, bone system, and peritoneum. The species most frequently reported was C. immitis. The most used treatment was azoles, followed by their combination with amphotericin B, monotherapy with amphotericin B, and alternative medicine. This work shows that epidemiological data outside the USA are still scarce. Serological tests are the preferred diagnostic method in daily medical practice, and cultures remain the gold standard. The treatment for coccidioidomycosis is ketoconazole and amphotericin B, individually or in combination.

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

Cited by 2 publications

References 31 publications

A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis

A multicentre external quality assessment: A first step to standardise PCR protocols for the diagnosis of histoplasmosis and coccidioidomycosis

Clinical Cases of Coccidioidomycosis in the Americas in the Period 1950–2021: Epidemiology Data, Diagnosis, and Treatment

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