Molecular probes for the detection of pathogenic fungi in the presence of human tissue

Kappe, Reinhard; Okeke, C. N.; Fauser, Christina; Maiwald, Matthias; Sonntag, Hans‐Günther

doi:10.1099/00222615-47-9-811

Cited by 67 publications

(58 citation statements)

References 23 publications

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“…PCR assays have been reported to increase sensitivity and reliability of fungal detection (Hendolin et al, 2000;Catten et al, 2001). However, results obtained by panfungal DNA amplification techniques with or without genus-specific probes (Einsele et al, 1997;Kappe et al, 1998) differ from those obtained with genus-specific primers (Skladny et al, 1999). The exact value of PCR for fungus detection in patients suffering from CRS has not yet been evaluated, particularly in NLs pretreated with a mucolytic agent.…”

Section: Introductionmentioning

confidence: 78%

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

Polzehl

Weschta

Podbielski

et al. 2005

Journal of Medical Microbiology

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Chronic rhinosinusitis (CRS) affects approximately 15 % of the adult population in industrialized countries. Fungi have been recognized as important pathogens in CRS in the immunocompromised host. Recently, fungi have been detected in more than 90 % of nasal lavages (NLs) in immunocompetent patients with CRS. Employing NLs of immunocompetent patients with CRS in the present study, the detection rates for fungi by culture techniques were compared with the results of different fungus-specific PCR assays. Standard fungal cultures were performed on NLs from 77 patients with CRS. NLs were also tested for the presence of fungal DNA by a panfungal assay with and without specific probes for Candida spp. and Aspergillus spp./Penicillium spp., and an Aspergillus-specific nested PCR assay. Nineteen of the 77 samples (25 %) grew fungi. Fungusspecific DNA was detected in 34 of 77 NLs (44 %). Twelve samples were positive for both culture and panfungal PCR, whereas seven specimens grew fungi in culture, but were negative in panfungal PCR, and an additional seven samples were positive in panfungal PCR, but negative in culture. The combination of culture and all employed PCR assays detected fungi in 39 patients (50 %). This study demonstrated that PCR and conventional culture techniques could be complementary diagnostic techniques to detect fungi in nasal specimens from CRS patients.

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Section: Introductionmentioning

confidence: 78%

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

Polzehl

Weschta

Podbielski

et al. 2005

Journal of Medical Microbiology

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“…However, the development of new techniques may enable a more accurate diagnosis than the diagnosis obtained with conventional methods [21]. Culture is still the method most used and many published studies do not report the identification of species and some noted a discrepancy in the determination of genus based on morphology and sequencing [19].…”

Section: Discussionmentioning

confidence: 99%

<i>Rhizopus oryzae</i> Retro-Orbital Abscess: Comparison of Spores and Hyphae Antifungal Susceptibility and Clinical Outcome

Fonseca¹,

Teixeira-Loyola²,

Lyra³

et al. 2018

OJMM

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Rhizopus is fungus of the order Mucorales, responsible for invasive diseases called mucormycosis. The correct identification of these fungi is one of the main questions raised in the literature since the diagnosis is not simple, being the necessary detection in biopsies of infected tissues and isolation of the microorganism for identification by morphological and molecular methods. In vitro susceptibility testing results are not fully clear since there are reports showing that in vitro resistance does not always mean therapeutic failure. This paper describes a case of rhino-orbito-cerebral mucormycosis in a patient with alcoholic chronic pancreatitis and secondary mellitus diabetes involving microbiological diagnostic, antifungal susceptibility tests for conidia and hypha and clinical correlation. The causative agent was identified by morphological characteristics and DNA sequencing. Minimal Inhibitory Concentration (MIC) for amphotericin B (AMB), itraconazole (ITZ) and terbinafine (TERB) was determined by broth microdilution method. Standardization of the dynamic growth evaluation patterns using the automated system BCT (BioCell-Tracer®) was obtained allowing to determining hyphae growth control and evaluating the effect of different concentrations of amphotericin B, itraconazole and terbinafine directly on growing hyphae. The fractional inhibitory concentration (FIC) for the combination of the two antifungals that the patient received at the same time, terbinafine and itraconazole, was also determined trying to correlate with clinical outcome. Strains isolated of retro-orbital abscess were identified as Rhizopus oryzae. MIC results (MIC) testing conidia showed high sensibility to amphotericin B and itraconazole in comparison to the obtained for terbinafine. The tests in the automated system BCT using hyphae showed sensibility to all tested antifungal agents and the FIC results for combination in vitro between terbinafine and itraconazole showed synergism consistent with the therapeutic success obtained for the patient who received initial treatment with amphotericin B followed for maintenance treatment with the combination itraconazole and terbinafine

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“…Dentre eles, citamos: uso de marcadores moleculares para identificação de fungos nos tecidos humanos (50) , a reação da cadeia de polimerase (10) , dosagens séricas e vítreas de Darabinitol, beta-D-glucan e do antígeno de C. albicans (51) , uso de anticorpos monoclonais radioiodinizados (52) e dosagem de anticorpos IgG no humor aquoso (53) . Os novos métodos descritos acima abrem novas perspectivas diagnósticas, porém necessitam de maiores estudos.…”

Section: Métodos Diagnósticosunclassified

Endoftalmite por Candida albicans

Serracarbassa¹,

Dotto²

2003

Arq. Bras. Oftalmol.

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Endoftalmite por Candida albicansO autor descreve os aspectos epidemiológicos, histopatológicos e clínicos da endoftalmite endógena por Candida albicans. Apresenta ainda novos métodos diagnósticos e opções terapêuticas utilizadas no tratamento das infecções fúngicas intra-oculares, por meio de revisão bibliográfica. INTRODUÇÃOOs fungos são os agentes mais comuns na endoftalmite endógena, representados na sua maioria pelas espécies de Candida.Infecções por Candida têm sido observadas desde os tempos de Hipó-crates, que descreveu a doença em pacientes debilitados.A endoftalmite resultante da infecção sistêmica por Candida foi inicialmente descrita em um olho enucleado em 1943 (1) . Em 1958, foi relatado o primeiro caso clinicamente reconhecido como endoftalmite endógena por Candida, confirmado posteriormente por necropsia (2) . Até a metade do século vinte, poucos casos de endoftalmite fúngica endógena foram relatados, porém, na década de sessenta, um grande núme-ro de trabalhos foi publicado a respeito, fato este que coincidiu com a introdução dos antibióticos de largo espectro (3) . Os fatores que contribuí-ram para o aumento da incidência de endoftalmite fúngica endógena nas últimas décadas incluem a difusão da antibioticoterapia, hiperalimentação intravenosa, terapia imunossupressiva, além do aumento da sobrevida dos pacientes imunocomprometidos, inclusive aqueles portadores da síndrome da imunodeficiência adquirida (AIDS) (4) . Após 1975, foram estudados um total de 167 casos descritos em 52 diferentes publicações (3) . Aproximadamente um terço dos casos teve envolvimento bilateral. As condições predisponentes foram similares àquelas relatadas antes de 1975, porém, associações com usuários de drogas ilícitas, prematuridade e AIDS são mais comumente encontradas nos relatos atuais (5) . As características dos achados oculares combinados com os achados clínicos e cultura positiva para Candida extra-ocular foram consideradas diagnósticas na maioria dos casos. Pelas descrições, as espécies de Candida, principalmente a C. albicans, foram as principais responsáveis por estas infecções fúngicas.A C. albicans é um habitante normal do trato gastrointestinal e regiões mucocutâneas, incluindo boca e vagina. Por ser um organismo que habita normalmente o corpo humano, culturas de sangue positivas são freqüente-mente atribuídas à contaminação, o que pode levar a um atraso no diagnós-tico de uma infecção por Candida.Imunossupressão, debilidade orgânica, entre outros fatores, podem tornar o fungo patogênico, resultando em uma grave infecção sistêmica (6) . O uso de drogas endovenosas ilícitas é a causa mais comum de endof-

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Molecular probes for the detection of pathogenic fungi in the presence of human tissue

Cited by 67 publications

References 23 publications

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

<i>Rhizopus oryzae</i> Retro-Orbital Abscess: Comparison of Spores and Hyphae Antifungal Susceptibility and Clinical Outcome

Endoftalmite por Candida albicans

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Molecular probes for the detection of pathogenic fungi in the presence of human tissue

Cited by 67 publications

References 23 publications

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

Fungus culture and PCR in nasal lavage samples of patients with chronic rhinosinusitis

&lt;i&gt;Rhizopus oryzae&lt;/i&gt; Retro-Orbital Abscess: Comparison of Spores and Hyphae Antifungal Susceptibility and Clinical Outcome

Endoftalmite por Candida albicans

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<i>Rhizopus oryzae</i> Retro-Orbital Abscess: Comparison of Spores and Hyphae Antifungal Susceptibility and Clinical Outcome