PCR Followed by Electrospray Ionization Mass Spectrometry for Broad-Range Identification of Fungal Pathogens

Massire, Christian; Buelow, Daelynn R.; Zhang, Sean X.; Lovari, Robert; Matthews, Heather; Toleno, Donna M.; Ranken, Raymond; Hall, Thomas A.; Metzgar, David; Sampath, Rangarajan; Blyn, Lawrence B.; Ecker, David J.; Gu, Zheng; Walsh, Thomas J.; Hayden, Randall T.

doi:10.1128/jcm.02621-12

Cited by 45 publications

(31 citation statements)

References 36 publications

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“…While our IFI case definition relies on culture and histopathology results, molecular diagnostic assays (i.e., polymerase chain reaction-based) also exist for fungal identification [21, 22]. However, they are not readily available at most clinical laboratories and, therefore, not yet an effective tool for early diagnosis.…”

Section: Discussionmentioning

confidence: 99%

Combat trauma-associated invasive fungal wound infections: epidemiology and clinical classification

et al. 2014

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Summary The emergence of invasive fungal wound infections (IFI) among combat casualties led to development of a combat trauma-specific IFI case definition and classification. Prospective data were collected from 1133 United States military personnel injured in Afghanistan (June 2009 through August 2011). The IFI rates ranged from 0.2% to 11.7% among ward and intensive care unit admissions, respectively (6.8% overall). Seventy-seven IFI cases were classified as proven/probable (n=54) and possible/unclassifiable (n=23) and compared in a case-case analysis. There was no difference in clinical characteristics between the proven/probable and possible/unclassifiable cases. Possible IFI cases had shorter time to diagnosis (p=0.02) and initiation of antifungal therapy (p=0.05) and fewer operative visits (p=0.002) compared to proven/probable cases, but clinical outcomes were similar between the groups. Although the trauma-related IFI classification scheme did not provide prognostic information, it is an effective tool for clinical and epidemiological surveillance and research.

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Section: Discussionmentioning

confidence: 99%

Combat trauma-associated invasive fungal wound infections: epidemiology and clinical classification

et al. 2014

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“…It is therefore evident that given the indisputable and ever-growing evidence of its superior, easier, and faster performance, MALDI-TOF MS has the potential to essentially replace conventional methods for identification of fungal pathogens in the next few years. Note that an alternative technique that has the same physical basis is the coupling of PCR amplification with electrospray ionization mass spectrometry (PCR-ESI MS) (282). However, due to the short period since its first development, data from clinical studies are still too immature to reach a valid conclusion about its clinical relevance (283).…”

Section: Novel Molecular Methodsmentioning

confidence: 99%

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

et al. 2014

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SUMMARY Invasive fungal infections constitute a serious threat to an ever-growing population of immunocompromised individuals and other individuals at risk. Traditional diagnostic methods, such as histopathology and culture, which are still considered the gold standards, have low sensitivity, which underscores the need for the development of new means of detecting fungal infectious agents. Indeed, novel serologic and molecular techniques have been developed and are currently under clinical evaluation. Tests like the galactomannan antigen test for aspergillosis and the β-glucan test for invasive Candida spp. and molds, as well as other antigen and antibody tests, for Cryptococcus spp., Pneumocystis spp., and dimorphic fungi, have already been established as important diagnostic approaches and are implemented in routine clinical practice. On the other hand, PCR and other molecular approaches, such as matrix-assisted laser desorption ionization (MALDI) and fluorescence in situ hybridization (FISH), have proved promising in clinical trials but still need to undergo standardization before their clinical use can become widespread. The purpose of this review is to highlight the different diagnostic approaches that are currently utilized or under development for invasive fungal infections and to identify their performance characteristics and the challenges associated with their use.

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“…As for mycological examination, it requires skill and expertise and may lead to false-negative results for polymicrobial specimens like sputum samples because of the more rapid and more extensive growth of other molds frequently associated with this fungus, like Aspergillus fumigatus (19). Several molecular methods have been proposed for detection of the fungus from sputum samples (20)(21)(22)(23)(24), but as culture methods, they do not allow the differentiation between airway colonization and sensitization of the patient or respiratory infection in the CF context, which has important implications for patient management.…”

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confidence: 99%

Purification and Characterization of a Mycelial Catalase from Scedosporium boydii, a Useful Tool for Specific Antibody Detection in Patients with Cystic Fibrosis

Mina¹,

Marot-Leblond²,

Cimon³

et al. 2014

Clin. Vaccine Immunol.

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bScedosporium boydii is an opportunistic filamentous fungus which may be responsible for a wide variety of infections in immunocompetent and immunocompromised individuals. This fungus belongs to the Scedosporium apiospermum species complex, which usually ranks second among the filamentous fungi colonizing the airways of patients with cystic fibrosis (CF) and may lead to allergic bronchopulmonary mycoses, sensitization, or respiratory infections. Upon microbial infection, host phagocytic cells release reactive oxygen species (ROS), such as hydrogen peroxide, as part of the antimicrobial response. Catalases are known to protect pathogens against ROS by detoxification of the hydrogen peroxide. Here, we investigated the catalase equipment of Scedosporium boydii, one of the major pathogenic species in the S. apiospermum species complex. Three catalases were identified, and the mycelial catalase A1 was purified to homogeneity by a three-step chromatographic process. This enzyme is a monofunctional tetrameric protein of 460 kDa, consisting of four 82-kDa glycosylated subunits. The potential usefulness of this enzyme in serodiagnosis of S. apiospermum infections was then investigated by an enzyme-linked immunosorbent assay (ELISA), using 64 serum samples from CF patients. Whatever the species involved in the S. apiospermum complex, sera from infected patients were clearly differentiated from sera from patients with an Aspergillus fumigatus infection or those from CF patients without clinical and biological signs of a fungal infection and without any fungus recovered from sputum samples. These results suggest that catalase A1 is a good candidate for the development of an immunoassay for serodiagnosis of infections caused by the S. apiospermum complex in patients with CF. Scedosporium boydii (formerly known as Pseudallescheria boydii) is one of the major pathogenic species within the Scedosporium apiospermum complex, which comprises four other species, namely, Scedosporium apiospermum sensu stricto, Scedosporium aurantiacum, Scedosporium minutisporum, and Scedosporium dehoogii, Scedosporium prolificans having been reassigned recently to the genus Lomentospora (Lomentospora prolificans) (1-4). These filamentous fungi are soilborne fungi that may cause a wide range of infections in humans, including subcutaneous mycetomas and ocular, bone, or joint infections resulting from traumatic inoculation of some fungal elements and infections of the respiratory tract (i.e., sinusitis and lung fungus ball), which are thought to be due to the inhalation of some airborne conidia (5-7). However, these fungi have gained attention during the past 2 decades mainly because of their recognition as common agents of colonization of the airways in patients with cystic fibrosis (CF). When appropriate culture media are used, the S. apiospermum species complex ranks second among the filamentous fungi recovered from respiratory specimens, with a prevalence ranging from 4.5 to 11.6% in patients (8-12). Although usually asymptomatic, this fungal colo...

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PCR Followed by Electrospray Ionization Mass Spectrometry for Broad-Range Identification of Fungal Pathogens

Cited by 45 publications

References 36 publications

Combat trauma-associated invasive fungal wound infections: epidemiology and clinical classification

Combat trauma-associated invasive fungal wound infections: epidemiology and clinical classification

Molecular and Nonmolecular Diagnostic Methods for Invasive Fungal Infections

Purification and Characterization of a Mycelial Catalase from Scedosporium boydii, a Useful Tool for Specific Antibody Detection in Patients with Cystic Fibrosis

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