magplex-tag microspheres  (DiaSorin Biotechnology)

Journal: PLoS ONE

Article Title: Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

doi: 10.1371/journal.pone.0084743

Figure Lengend Snippet: Predictions of the number of isolates of each pospiviroid species that will be detected using universal and species-specific assays of the Luminex MagPlex-TAG pospiviroid array.

Article Snippet: To prepare the mixture of 11 optically distinct Luminex MagPlex-TAG beads , each bottle of MagPlex-TAG microspheres was vortexed vigorously for 1 min, then sonicated for 30 s. Bead mixtures were prepared in Eppendorf LoBind tubes (Eppendorf, Hamburg, Germany), diluted with 2×Tm hybridization buffer (0.2 M Tris-HCl pH 8.0, 0.4 M NaCl, 0.16% Triton X-100) to yield a final concentration of approximately 1250 beads of each bead address per 25 µL.

Techniques: Luminex

Journal: PLoS ONE

Article Title: Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

doi: 10.1371/journal.pone.0084743

Figure Lengend Snippet: Flow diagram depicting the five steps of the Luminex MagPlex-TAG pospiviroid array method.

Article Snippet: To prepare the mixture of 11 optically distinct Luminex MagPlex-TAG beads , each bottle of MagPlex-TAG microspheres was vortexed vigorously for 1 min, then sonicated for 30 s. Bead mixtures were prepared in Eppendorf LoBind tubes (Eppendorf, Hamburg, Germany), diluted with 2×Tm hybridization buffer (0.2 M Tris-HCl pH 8.0, 0.4 M NaCl, 0.16% Triton X-100) to yield a final concentration of approximately 1250 beads of each bead address per 25 µL.

Techniques: Luminex

Journal: PLoS ONE

Article Title: Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

doi: 10.1371/journal.pone.0084743

Figure Lengend Snippet: The sensitivity of the multiplexed Luminex MagPlex-TAG pospiviroid array when tested using a ten-fold serial dilutions of a total RNA extract from a Solanum lycopersicum sample infected with Potato spindle tuber viroid (PSTVd, isolate #N).

Article Snippet: To prepare the mixture of 11 optically distinct Luminex MagPlex-TAG beads , each bottle of MagPlex-TAG microspheres was vortexed vigorously for 1 min, then sonicated for 30 s. Bead mixtures were prepared in Eppendorf LoBind tubes (Eppendorf, Hamburg, Germany), diluted with 2×Tm hybridization buffer (0.2 M Tris-HCl pH 8.0, 0.4 M NaCl, 0.16% Triton X-100) to yield a final concentration of approximately 1250 beads of each bead address per 25 µL.

Techniques: Luminex, Infection

Journal: PLoS ONE

Article Title: Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

doi: 10.1371/journal.pone.0084743

Figure Lengend Snippet: Percentages of false positive and false negative results for Luminex MagPlex-TAG pospiviroid array data when analyzed using three different methods for setting thresholds for positivity.

Article Snippet: To prepare the mixture of 11 optically distinct Luminex MagPlex-TAG beads , each bottle of MagPlex-TAG microspheres was vortexed vigorously for 1 min, then sonicated for 30 s. Bead mixtures were prepared in Eppendorf LoBind tubes (Eppendorf, Hamburg, Germany), diluted with 2×Tm hybridization buffer (0.2 M Tris-HCl pH 8.0, 0.4 M NaCl, 0.16% Triton X-100) to yield a final concentration of approximately 1250 beads of each bead address per 25 µL.

Techniques: Luminex

Journal: PLoS ONE

Article Title: Development of a Multiplexed Bead-Based Suspension Array for the Detection and Discrimination of Pospiviroid Plant Pathogens

doi: 10.1371/journal.pone.0084743

Figure Lengend Snippet: Details of pospiviroid isolates used for validation of the Luminex MagPlex-TAG pospiviroid array.

Article Snippet: To prepare the mixture of 11 optically distinct Luminex MagPlex-TAG beads , each bottle of MagPlex-TAG microspheres was vortexed vigorously for 1 min, then sonicated for 30 s. Bead mixtures were prepared in Eppendorf LoBind tubes (Eppendorf, Hamburg, Germany), diluted with 2×Tm hybridization buffer (0.2 M Tris-HCl pH 8.0, 0.4 M NaCl, 0.16% Triton X-100) to yield a final concentration of approximately 1250 beads of each bead address per 25 µL.

Techniques: Luminex

Journal: Laboratory Medicine

Article Title: Multiplex Microsphere PCR (mmPCR) Allows Simultaneous Gram Typing, Detection of Fungal DNA, and Antibiotic Resistance Genes

doi: 10.1093/labmed/lmac023

Figure Lengend Snippet: Simultaneous detection of Gram type, fungi, antibiotic resistance genes, and species-specific genes with multiplex microsphere polymerase chain reaction (mmPCR). The signal-to-noise ratio (S/N) of (13×) 16S/rRNA Sanger-sequenced and Basic Local Alignment Tool-identified bacterial isolates and (3×) fungal isolates, selectively amplified in a custom 10-plex mmPCR assay. Each 10-plex mmPCR contained general Gram-positive (G-pos), Gram-negative (G-neg) and pan-fungal (Fungi) primers; specific primers targeting the resistance-conferring gene mecA (MecA), the type A vancomycin resistance–conferring Tn1546 Transposon (VanA), the vanB mobile cluster (VanB), and the β-lactamases expressing bla SHV-1 gene (β-lac); and specific primers targeting the bacterial species B cepacia (B.cep), P aeruginosa (P.aer), and A xylosoxidans (A.xyl). The polymerase chain reaction was conducted using purified isolate genomic DNA at a concentration of 10 5 genomes/reaction. The S/N of each primer set for each isolate was calculated from data normalized to a no-template negative control reaction (noise) and a “signal maximum” reaction that contained no template or forward and reverse primers (signal). An S/N >0.0 was considered positive and weakly positive when between 0.0 and 0.2 (dotted line). An S/N less than –0.1 was omitted. Data are representative of triplicate independent technical replicates. The mean ± standard error of the mean of technical triplicates is shown.

Article Snippet: Isolated gDNA of 16S/rRNA Sanger-sequenced and Basic Local Alignment Tool–identified bacterial and fungal isolates were selectively amplified in a custom 10-plex Luminex MagPlex-TAG microsphere-based mmPCR assay.

Techniques: Multiplex Assay, Polymerase Chain Reaction, Amplification, Expressing, Purification, Concentration Assay, Negative Control