An automated PCR platform with homogeneous time-resolved fluorescence detection and dry chemistry assay kits

Hagren, Virve; Lode, Piia von; Syrjälä, Anniina; Soukka, Tero; Lövgren, Timo; Kojola, Hannu; Nurmi, Johanna

doi:10.1016/j.ab.2007.12.017

Cited by 16 publications

(21 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Each chip also contains an internal amplification control of a non-naturally existing DNA sequence, which is used to monitor for the presence of inhibitors and the integrity of the dried reagents. The metallic background of the chips ensures efficient transfer of heat from the thermocycler to the PCR reaction and thus decreases the time required for the target amplification [62].…”

Section: Assay Methodology and Automation Pcr Test Chipmentioning

confidence: 99%

“…The assays take place in disposable plastic reaction vessels, in other words the 'test chips'. The test chips readily contain all the assay-specific PCR reagents, including the polymerase, in dry form [62]. Each chip also contains an internal amplification control of a non-naturally existing DNA sequence, which is used to monitor for the presence of inhibitors and the integrity of the dried reagents.…”

Section: Assay Methodology and Automation Pcr Test Chipmentioning

confidence: 99%

“…In the GenomEra format, the detection of the amplification products is based on the company's proprietary technique called enhanced competitive hybridization where two partially complementary oligonucleotide probes are used [62]. One of the probes (the label probe) is labeled with a time-resolved fluorescent label, terbium (Tb) for the mecA target and europium (Eu) for the S. aureus target, while the other probe (the quencher probe) is labeled with a quencher molecule.…”

Section: Probe Technologymentioning

confidence: 99%

“…The main functions of the GenomEra CDX instrument include the detection and identification of inserted test chips via the barcodes on the chips, the irreversible sealing of the test chips using heat and pressure to minimize the risk of cross-contamination, thermocycling (i.e., nucleic acid amplification) by moving the chips between a range of heated blocks at different temperatures, and homogeneous measurement of fluorescence directly from the chips in two different temperatures at the end of the assay run [62]. The entire assay sequence from the beginning of the sample preparation to the reporting of results takes less than an hour [63,64].…”

Section: Thermal Cyclingmentioning

confidence: 99%

See 3 more Smart Citations

GenomEra MRSA/SA, a fully automated homogeneous PCR assay for rapid detection ofStaphylococcus aureusand the marker of methicillin resistance in various sample matrixes

Hirvonen

Kaukoranta

2013

Expert Review of Molecular Diagnostics

View full text Add to dashboard Cite

The GenomEra MRSA/SA assay (Abacus Diagnostica, Turku, Finland) is the first commercial homogeneous PCR assay using thermally stable, intrinsically fluorescent time-resolved fluorometric (TRF) labels resistant to autofluorescence and other background effects. This fully automated closed tube PCR assay simultaneously detects Staphylococcus aureus specific DNA and the mecA gene within 50 min. It can be used for both screening and confirmation of methicillin-resistant and -sensitive S. aureus (MRSA and MSSA) directly in different specimen types or from preceding cultures. The assay has shown excellent performance in comparisons with other diagnostic methods in all the sample types tested. The GenomEra MRSA/SA assay provides rapid assistance for the detection of MRSA as well as invasive staphylococcal infections and helps the early targeting of antimicrobial therapy to patients with potential MRSA infection.

show abstract

Section: Assay Methodology and Automation Pcr Test Chipmentioning

confidence: 99%

Section: Assay Methodology and Automation Pcr Test Chipmentioning

confidence: 99%

Section: Probe Technologymentioning

confidence: 99%

Section: Thermal Cyclingmentioning

confidence: 99%

See 2 more Smart Citations

GenomEra MRSA/SA, a fully automated homogeneous PCR assay for rapid detection ofStaphylococcus aureusand the marker of methicillin resistance in various sample matrixes

Hirvonen

Kaukoranta

2013

Expert Review of Molecular Diagnostics

View full text Add to dashboard Cite

show abstract

“…Since the signal-to-noise ratios of either the HLA-B*27-probe or the amplification control probe did not show any clear decline during this period it could be assumed that even longer storage time could be used, and these tests are continuing in our laboratory. Storing the reagents in lower temperatures, such as at +4 • C or even at −20 • C, would likely result in even longer self-life than expected for storage at room temperature [5,6].…”

Section: Discussionmentioning

confidence: 99%

A Homogeneous HLA-B*27 Genotyping Assay Using Dried Reagent Mixtures

Ilonen¹,

Lövgren

2009

Disease Markers

Self Cite

View full text Add to dashboard Cite

Abstract. The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance.

show abstract

The microbial threat: Can rare earths help?

Lezhnina

Rochowiak

Göhde

et al. 2020

Journal of Biophotonics

View full text Add to dashboard Cite

Despite an ever increasing demand for reliable and cheap methods in the detection and quantification of microbes, surprisingly few investigations have explored or utilized the luminescence of rare earths in the microbial context, neither in conventional, that is, plating and microscopic imaging techniques, nor in advanced methods like fluorescence flow cytometry. We have thus investigated the potential of some rare earth complexes and hybrid materials for microbiological analysis. We found fairly simple procedures for internal staining (dyes inside the bacterial cell) and external staining (dyes on the cell surface). The present paper is predominantly relying on microscopic imaging and luminescence spectroscopies (excitation, emission, decay times), but also evaluates model rare earth microspheres to estimate an eventual rare earth based stain for a fast and sensitive bacteria enumeration with luminescence flow cytometry.

show abstract

An automated PCR platform with homogeneous time-resolved fluorescence detection and dry chemistry assay kits

Cited by 16 publications

References 25 publications

GenomEra MRSA/SA, a fully automated homogeneous PCR assay for rapid detection ofStaphylococcus aureusand the marker of methicillin resistance in various sample matrixes

GenomEra MRSA/SA, a fully automated homogeneous PCR assay for rapid detection ofStaphylococcus aureusand the marker of methicillin resistance in various sample matrixes

A Homogeneous HLA-B*27 Genotyping Assay Using Dried Reagent Mixtures

The microbial threat: Can rare earths help?

Contact Info

Product

Resources

About