Molecular diagnostic methods for invasive fungal disease: the horizon draws nearer?

Halliday, Catriona; Kidd, Sarah; Sorrell, Tania C.; Chen, S.C.-A.

doi:10.1097/pat.0000000000000234

Cited by 46 publications

(21 citation statements)

References 156 publications

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“…Currently culture, and other conventional, diagnostic methods are still the cornerstone of diagnosis. Molecular methods to directly detect/identify mould pathogens in tissue and body fluids are increasingly used [164].…”

Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

“…Nucleic acid amplification techniques that employ either panfungal PCR assays (combined with DNA sequencing) or fungal species-specific assays may also be used to identify pathogens directly in tissue, blood, serum or sputum [164]. These methods yielded good sensitivity and specificity in small case series [3, 167,168] but methods are not standardized.…”

Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

“…Apophysomyces and other genera within the Mucorales, and genera, including Fusarium and Scedosporium, species complexes. [164,173,174]. Fungal identification by molecular methods is largely reliant on DNA sequencing and comparative sequence analysis is now the reference standard for identification of filamentous fungi.…”

Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

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Emerging infections caused by non- Aspergillus filamentous fungi

Douglas

Chen

Slavin

2016

Clinical Microbiology and Infection

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There are three broad groups of non-Aspergillus moulds: the mucormycetes, the hyalohyphomycetes and the phaeohyphomycetes. Infections with these pathogens are increasingly reported, particularly in the context of increasing use of immunosuppressant agents and improved diagnostics. The epidemiology of non-Aspergillus mould infections varies with geography, climate and level of immunosuppression. Skin and soft-tissue infections are the predominant presentation in the immunocompetent host and pulmonary and other invasive infections in the immunocompromised host. The more common non-Aspergillus moulds include Rhizopus, Mucor, Fusarium and Scedosporium species; however, other emerging pathogens are Rasamsonia and Verruconis species, which are discussed in this article. Outbreaks of non-Aspergillus mould infections have been increasingly reported, with contaminated medical supplies and natural disasters as common sources. Currently culture and other conventional diagnostic methods are the cornerstone of diagnosis. Molecular methods to directly detect and identify mould pathogens in tissue and body fluids are increasingly used.

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Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

Section: Advances In Laboratory Diagnosticsmentioning

confidence: 99%

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Emerging infections caused by non- Aspergillus filamentous fungi

Douglas

Chen

Slavin

2016

Clinical Microbiology and Infection

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“…The usefulness of molecular methods based on PCR for the proper detection of fungal species in IFIs has been proven (25,26). However, several limitations exist when the fungus causing the infection is unknown.…”

Section: Discussionmentioning

confidence: 99%

New Panfungal Real-Time PCR Assay for Diagnosis of Invasive Fungal Infections

et al. 2016

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The diagnosis of invasive fungal infections (IFIs) is usually based on the isolation of the fungus in culture and histopathological techniques. However, these methods have many limitations often delaying the definitive diagnosis. In recent years, molecular diagnostics methods have emerged as a suitable alternative for IFI diagnosis. When there is not a clear suspicion of the fungus involved in the IFI, panfungal real-time PCR assays have been used, allowing amplification of any fungal DNA. However, this approach requires subsequent amplicon sequencing to identify the fungal species involved, increasing response time. In this work, a new panfungal real-time PCR assay using the combination of an intercalating dye and sequence-specific probes was developed. After DNA amplification, a melting curve analysis was also performed. The technique was standardized by using 11 different fungal species and validated in 60 clinical samples from patients with proven and probable IFI. A melting curve database was constructed by collecting those melting curves obtained from fungal species included in the standardization assay. Results showed high reproducibility (coefficient of variation [CV] < 5%; r > 0.95) and specificity (100%). The overall sensitivity of the technique was 83.3%, with the group of fungi involved in the infection detected in 77.8% of the positive samples with IFIs covered by molecular beacon probes. Moreover, sequencing was avoided in 67.8% of these "probe-positive" results, enabling report of a positive result in 24 h. This technique is fast, sensitive, and specific and promises to be useful for improving early diagnosis of IFIs.

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“…Son técnicas que pueden aplicarse directamente a la muestra clínica o a la colonia obtenida del cultivo. Numerosos artículos han revisado la situación actual de estas nuevas técnicas de diagnóstico molecular frente a otras técnicas de diagnóstico clásico [15][16][17] .…”

Section: Detección De Hongosunclassified

Métodos moleculares de diagnóstico de infecciones respiratorias. ¿Ha cambiado el esquema diagnóstico?

Vila

Zboromyrska

Vergara

et al. 2016

Enfermedades Infecciosas y Microbiología Clínica

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Lower respiratory tract infections remain one of the most common causes of mortality worldwide, which is why early diagnosis is crucial. Traditionally the microbiological diagnosis of these infections has been based on conventional methods including culture on artificial media for isolation of bacteria and fungi and cell cultures for virus and antibody or antigen detection using antigen-antibody reactions. The main drawback of the above mentioned methods is the time needed for an etiological diagnosis of the infection. The techniques based on molecular biology have drawn much attention in recent decades as tools for rapid diagnosis of infections. Some techniques are very expensive, especially those that can detect various microorganisms in the same reaction, therefore the question that arises is whether the cost of such testing is justified by the information obtained and by the clinical impact that its implementation will determine. In this article we make a review of the various techniques of molecular biology applied to the diagnosis of pneumonia and focus primarily on analysing the impact they may have on the management of patients with acute respiratory tract infections.

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Molecular diagnostic methods for invasive fungal disease: the horizon draws nearer?

Cited by 46 publications

References 156 publications

Emerging infections caused by non- Aspergillus filamentous fungi

Emerging infections caused by non- Aspergillus filamentous fungi

New Panfungal Real-Time PCR Assay for Diagnosis of Invasive Fungal Infections

Métodos moleculares de diagnóstico de infecciones respiratorias. ¿Ha cambiado el esquema diagnóstico?

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