Use of a Panfungal PCR Assay for Detection of Fungal Pathogens in a Commercial Blood Culture System

Iwen, Peter C.; Freifeld, Alison G.; Bruening, Tricia A.; Hinrichs, Steven H.

doi:10.1128/jcm.42.5.2292-2293.2004

Cited by 20 publications

(10 citation statements)

References 20 publications

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“…This has been achieved using PCR product sequencing (9,11,14), although this remains relatively costly, time-consuming, and potentially inaccurate in the presence of mixed fungal species. Other methods, such as differences in PCR product sizes following electrophoresis, restriction fragment length polymorphism analysis, or single-stranded conformational polymorphism analysis (2, 13), have been used but are not sequence specific and may not be readily adapted for use in the clinical microbiology laboratory.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Detection and Identification of Candida , Aspergillus , and Cryptococcus Species by Reverse Line Blot Hybridization

et al. 2006

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We report on a reverse line blot (RLB) assay, utilizing fungal species-specific oligonucleotide probes to hybridize with internal transcribed spacer 2 region sequences amplified using a nested panfungal PCR. Reference and clinical strains of 16 Candida species (116 strains), Cryptococcus neoformans (five strains of Cryptococcus neoformans var. neoformans, five strains of Cryptococcus neoformans var. grubii, and six strains of Cryptococcus gatti), and five Aspergillus species (68 strains) were all correctly identified by the RLB assay. Additional fungal species (16 species and 26 strains) not represented on the assay did not exhibit crosshybridization with the oligonucleotide probes. In simulated clinical specimens, the sensitivity of the assay for Candida spp. and Aspergillus spp. was 10 0.5 cells/ml and 10 2 conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens.

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Section: Discussionmentioning

confidence: 99%

Simultaneous Detection and Identification of Candida , Aspergillus , and Cryptococcus Species by Reverse Line Blot Hybridization

et al. 2006

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“…The universal detection of bacteria or fungi in blood using broad-range PCR assays has been described (1,16,22,42), but interpretations can be difficult, as assay contaminants can cause false-positive results (37). The early detection of BSIs requires reliable sensitivity, around 10 CFU/ml of blood for bacteria and fungi, which is below the level of most existing diagnostic platforms.…”

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confidence: 99%

Rapid Real-Time Nucleic Acid Sequence-Based Amplification-Molecular Beacon Platform To Detect Fungal and Bacterial Bloodstream Infections

Zhao¹,

Park²,

Kreiswirth³

et al. 2009

J Clin Microbiol

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Bloodstream infections (BSIs) are a significant cause of morbidity and mortality. Successful patient outcomes are diminished by a failure to rapidly diagnose these infections and initiate appropriate therapy. A rapid and reliable diagnostic platform of high sensitivity is needed for the management of patients with BSIs. The combination of an RNA-dependent nucleic acid sequence-based amplification and molecular beacon (NASBA-MB) detection system in multiplex format was developed to rapidly detect medically important BSI organisms. Probes and primers representing pan-gram-negative, pan-gram-positive, pan-fungal, pan-Candida, and panAspergillus organisms were established utilizing 16S and 28S rRNA targets for bacteria and fungi, respectively. Two multiplex panels were developed to rapidly discriminate bacterial or fungal infections at the subkingdom/ genus level with a sensitivity of 1 to 50 genomes. A clinical study was performed to evaluate the accuracy of this platform by evaluating 570 clinical samples from a tertiary-care hospital group using blood bottle samples.

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“…Detection systems for metabolites (e.g., fungal D-arabinitol, (5,10), fungal DNA by PCR (1,8,11), and fungal cell wall components (e.g., ␤-D-glucan) (13,14) have all been investigated, but none has yet achieved broad validation. Assays for fungal antigens (or antibodies to same) have also been studied, but generally with little success (16).…”

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confidence: 99%

Syscan3, a Kit for Detection of Anti- Candida Antibodies for Diagnosis of Invasive Candidiasis

et al. 2005

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Use of a Panfungal PCR Assay for Detection of Fungal Pathogens in a Commercial Blood Culture System

Cited by 20 publications

References 20 publications

Simultaneous Detection and Identification of Candida , Aspergillus , and Cryptococcus Species by Reverse Line Blot Hybridization

Simultaneous Detection and Identification of Candida , Aspergillus , and Cryptococcus Species by Reverse Line Blot Hybridization

Rapid Real-Time Nucleic Acid Sequence-Based Amplification-Molecular Beacon Platform To Detect Fungal and Bacterial Bloodstream Infections

Syscan3, a Kit for Detection of Anti- Candida Antibodies for Diagnosis of Invasive Candidiasis

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