Development and evaluation of the nuclisens basic kit NASBA for the detection of RNA from Candida species frequently resistant to antifungal drugs

Loeffler, Juergen; Dorn, Christiane; Hebart, Holger; Cox, Philipp; Magga, Stella; Einsele, Hermann

doi:10.1016/s0732-8893(02)00510-2

Cited by 23 publications

(8 citation statements)

References 19 publications

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“…The detection limits of pan-Candida and pan-Aspergillus assays are in agreement with the results reported by other studies that used traditional NASBA detection (electrochemiluminescence) for Candida spp. (28,46) or Aspergillus spp. (29).…”

Section: Vol 47 2009 Molecular Beacon Detection Of Bloodstream Infementioning

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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Section: Vol 47 2009 Molecular Beacon Detection Of Bloodstream Infementioning

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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“…Previous reports on the application of "isothermal" nucleic acid amplification techniques to yeast identification (3,4,22,46) are all based on nucleic acid sequence-based amplification (6), but this method is rather unspecific due to the need to use a relatively low temperature (40°C) for amplification (33). We are interested in the development of a simple and user-friendly bench DNA-based diagnostic kit for the identification of clinically relevant yeasts.…”

mentioning

confidence: 99%

Efficient Identification of Clinically Relevant Candida Yeast Species by Use of an Assay Combining Panfungal Loop-Mediated Isothermal DNA Amplification with Hybridization to Species-Specific Oligonucleotide Probes

2008

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The occurrence of invasive mycoses has progressively increased in recent years. Yeasts of the genus Candida remain the leading etiologic agents of those infections. Early identification of opportunistic yeasts may contribute significantly to improved disease management and the selection of appropriate antifungal therapy. We developed a rapid and reliable molecular identification system for clinically relevant yeasts that makes use of nonspecific primers to amplify a region of the 26S rRNA gene, followed by reverse hybridization of the digoxigenin-labeled products to a panel of species-specific oligonucleotide probes arranged on a nylon membrane macroarray format. DNA amplification was achieved by the recently developed loop-mediated isothermal DNA amplification technology, a promising option for the development of improved laboratory diagnostic kits. The newly developed method was successful in distinguishing among the major clinically relevant yeasts associated with bloodstream infections by using simple, rapid, and cost-effective procedures and equipment.

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“…Amplification and detection can be completed within few hours and the analysis has shown a detection limit of 1 CFU. NASBA has been evaluated to detect six various Candida species [66]. The main benefits of NASBA compared to PCR are no need of thermal cycling instrument and specific detection of living yeast cells, because RNA unlike DNA is rapidly degraded outside cells.…”

Section: Nucleic Acid Sequence Based Amplification (Nasba)mentioning

confidence: 99%

Molecular-Genetic Approaches for Identification and Typing of Pathogenic Candida Yeasts: A Review

Singh¹

2014

IJIRSET

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Candida is a medically important fungi because of its high frequency as a commensal and pathogenic microorganism causing superficial as well as invasive infections. Because the accurate diagnosis of candidiasis remains difficult, a fast and reliable assay for characterization of fungal pathogens is critical for the early initiation of adequate antifungal therapy and/or for introduction of preventive measures. As novel molecular genetic techniques are continuously introduced, their role in the management of infectious diseases has also been growing .Strain typing and delineation of the species are essential for understanding its biology, epidemiology and population structure. A wide range of molecular techniques have been used for this purpose including non-DNA-based methods (multi-locus enzyme electrophoresis), conventional DNA-based methods (electrophoretic karyotyping, random amplified polymorphic DNA, amplified fragment length polymorphism, restriction enzyme analysis with and without hybridization, rep-PCR) and DNA-based methods called exact typing (PCR based methods, microstallite length polymorphism, multilocus sequence typing, DNA microarray ) because they generate unambiguous and highly reproducible typing data. Today, molecular strategies complement conventional methods and provide more accurate and detailed insight. It can be expected that future technical development will improve their potential furthermore.In this article, we provide a critical review on the vigorously fermenting field of molecular approaches to Candida identification and typing and summarized their advantages and limitations with regard to their discriminatory power, reproducibility, cost and ease of performance.

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Development and evaluation of the nuclisens basic kit NASBA for the detection of RNA from Candida species frequently resistant to antifungal drugs

Cited by 23 publications

References 19 publications

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

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

Efficient Identification of Clinically Relevant Candida Yeast Species by Use of an Assay Combining Panfungal Loop-Mediated Isothermal DNA Amplification with Hybridization to Species-Specific Oligonucleotide Probes

Molecular-Genetic Approaches for Identification and Typing of Pathogenic Candida Yeasts: A Review

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