Detection of Yeasts in Blood Cultures by the Luminex xTAG Fungal Assay

Balada-Llasat, Joan-Miquel; LaRue, Heidi; Kamboj, Kamal; Rigali, Lisa; Smith, Debra; Thomas, Keelie; Pancholi, Preeti

doi:10.1128/jcm.06375-11

Cited by 35 publications

(17 citation statements)

References 22 publications

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“…In recent years, microsphere-based technology has become an attractive alternative to DNA sequencing. This technology had been applied for species identification in molds and yeasts (16)(17)(18)(19)(20) and for the identification of single nucleotide polymorphisms (SNPs) (21,22). Here, we describe the adaptation of this technology to identify SNPs in the FKS1 HS1 and FKS2 HS1 domains of C. glabrata which confer in vitro resistance to echinocandins.…”

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confidence: 99%

Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

et al. 2014

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Echinocandins are the recommended treatment for invasive candidiasis due to Candida glabrata. Resistance to echinocandins is known to be caused by nonsynonymous mutations in the hot spot-1 (HS1) regions of the FKS1 and FKS2 genes, which encode a subunit of the ␤-1,3-glucan synthase, the target of echinocandins. Here, we describe the development of a microsphere-based assay using Luminex MagPix technology to identify mutations in the FKS1 HS1 and FKS2 HS1 domains, which confer in vitro echinocandin resistance in C. glabrata isolates. The assay is rapid and can be performed with high throughput. The assay was validated using 102 isolates that had FKS1 HS1 and FKS2 HS1 domains previously characterized by DNA sequencing. The assay was 100% concordant with DNA sequencing results. The assay was then used for high-throughput screening of 1,032 C. glabrata surveillance isolates. Sixteen new isolates with mutations, including a mutation that was new to our collection (del659F), were identified. This assay provides a rapid and cost-effective way to screen C. glabrata isolates for echinocandin resistance.

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Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

et al. 2014

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“…An 11-plex bead array was used by Buelow et al to identify common respiratory-associated fungi using the Luminex platform [54]. Babady et al used the same platform to design a Luminex-based assay to identify 23 fungi commonly found to cause invasive infections in the immunocompromised while Balada-Llasat et al used the Luminex platform to identify yeasts from blood cultures [55,56]. These assays are targeted approaches to identify anticipated groups of fungi.…”

Section: Arraysmentioning

confidence: 98%

Diagnostic Molecular Biology

Romanelli

Wickes

2015

Diagnosis and Treatment of Fungal Infections

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“…Flow cytometry is routinely used in the diagnosis of health disorders, especially blood cancers, but has many other applications in basic research and clinical practice. Especially the xMAP ® technology allows a high plex-grade for NAT-based target detection in microtitration plate scale [63]. Here, nucleic acids are extracted from target cells/several sample media and purified from the pretreated samples with commercially available kits.…”

Section: Flow Cytometrymentioning

confidence: 99%

Nucleic Acid Amplification Techniques

Lehmann¹,

Schmitz

2015

Modern Techniques for Pathogen Detection

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Nucleic acid amplification techniques (NATs), more commonly but imprecisely called polymerase chain reaction (PCR), have evolved, among others, as central tools for pathogen identification and strain typing. The huge field of applications of the technique in all its modifications already marks the centerpiece of diagnostic approaches and has not only opened the door to a completely new understanding of elementary and sophisticated metabolic processes but also is the basis of the most important trends in life science industries. In a sense, it acts as the lens of a molecular microscope which visualizes the code from the genomic book, while its meanings have to be deciphered by a further armory of laboratory avenues.In 1983, PCR was invented by Kary Mullis, for which he later (1993) was awarded with the Nobel Prize in Chemistry along with Michael Smith [1]. The protocol is based on targeting specific DNA or rDNA sequences with more or less specific short tracer sequences (primers/oligonucleotides, colloquially oligos) binding to flanking regions of the core sequence to be proliferated. A succession of thermal incubation steps defined by length and temperature are cycled and conduct double-stranded DNA (dsDNA) melting, that is, separation of the target DNA into antiparallel, complementary, single-stranded DNA (ssDNA) strands, specific binding of primers (annealing), and enzymatic amplification out of the basic set of four nucleotide monomers of the sequence embedded. In continuation, the newly synthesized dsDNA copies, selected in length by the specific primer binding sites (PBSs), themselves serve as templates for further replication rounds, setting in motion a chain reaction in which the DNA template is exponentially amplified. The core enzyme is a DNA polymerase, which was heat-sensitive at time of invention but subsequently was destroyed at the melting temperature. To gain useful numbers of target copies, an expensive and time-consuming replacement of the enzyme was needed at the beginning of each of the usually 20-40 replication cycles. The discovery of an extraordinarily thermostable DNA polymerase purified from the thermophilic bacterium Thermophilus aquaticus, which naturally lives in hot (50-80 ∘ C) environments such as hot springs [2], Modern Techniques for Pathogen Detection, First Edition. Edited by Jürgen Popp and Michael Bauer.

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Detection of Yeasts in Blood Cultures by the Luminex xTAG Fungal Assay

Cited by 35 publications

References 22 publications

Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

Development of a Luminex-Based Multiplex Assay for Detection of Mutations Conferring Resistance to Echinocandins in Candida glabrata

Diagnostic Molecular Biology

Nucleic Acid Amplification Techniques

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