| 1. |
- Rissanen, Maria, et al.
(författare)
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Novel homogenous time-resolved fluorometric RT-PCR assays for quantification of PSA and hK2 mRNAs in blood
- 2007
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Ingår i: Clinical Biochemistry. - : Elsevier BV. - 1873-2933 .- 0009-9120. ; 40:1-2, s. 111-118
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: The purpose of this study was to design, validate, and optimize internally standardized real-time quantitative RT-PCR assays and to identify and avoid problems with assay reliability and examine the impact of an exogenous internal standard. Design and methods: The model system consisted of internally standardized quantitative real-time RT-PCR assays specific for PSA and hK2 mRNA based on time-resolved fluorometric detection of lanthanide chelates. Results: Reproducibility was best when large copy numbers (> 5000 per milliliter blood) were analyzed. Addition of an exogenous target-mimicking internal standard had no significant effect on the reproducibility of the method, but increased the calculated copy numbers by an average of 2-fold. Conclusions: We developed an internally standardized, specific and reproducible real-time RT-PCR analysis method for PSA and hK2 mRNA in circulating cells in the bloodstream. Both PSA and hK2 assays are sufficiently sensitive to detect two LNCaP cells per milliliter whole blood. (c) 2006 The Canadian Society of Clinical Chemists. All rights reserved.
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| 2. |
- Daniel, Ondrej, et al.
(författare)
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Blind sub-Nyquist GNSS signal detection
- 2016
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Ingår i: ICASSP, IEEE International Conference on Acoustics, Speech and Signal Processing - Proceedings. - 1520-6149. ; 2016-May, s. 6575-6579
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Konferensbidrag (refereegranskat)abstract
- A satellite navigation receiver traditionally searches for positioning signals using an acquisition procedure. In situations, in which the required information is only a binary decision whether at least one positioning signal is present or absent, the procedure represents an unnecessarily complex solution. This paper presents a different approach for the binary detection problem with significantly reduced computational complexity. The approach is based on a novel decision metric which is utilized to design two binary detectors. The first detector operates under the theoretical assumption of additive white Gaussian noise and is evaluated by means of Receiver Operating Characteristics. The second one considers also additional interferences and is suitable to operate in a real environment. Its performance is verified using a signal captured by a receiver front-end.
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| 3. |
- Hepojoki, Satu, et al.
(författare)
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Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease
- 2015
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Ingår i: Journal of Clinical Microbiology. - 0095-1137 .- 1098-660X. ; 53:7, s. 2292-2297
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Tidskriftsartikel (refereegranskat)abstract
- In this study, we describe a competitive homogeneous immunoassay that makes use of Forster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease.
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| 4. |
- Nurmi, Jussi, et al.
(författare)
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Time-resolved fluorometry in end-point and real-time PCR quantification of nucleic acids
- 2000
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Ingår i: Luminescence. - 1522-7235. ; 15:6, s. 381-388
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Tidskriftsartikel (refereegranskat)abstract
- Two time-resolved fluorescence-based methods for nucleic acid quantification are described and their results are compared. Both methods use an exogenous internal standard to eliminate errors arising from different steps of the assay. The first method is a competitive end-point assay, where the standard competes for the same primers with the actual target sequence, prostate-specific antigen (PSA) cDNA. The standard and target are quantified in a dual-label plate hybridization with lanthanide-labelled probes after a fixed number of PCR cycles. The second method is based on real-time monitoring of PCR and on the use of a novel homogeneous signal generation principle that relies on the use of a 5′→3′ exonucleolytic DNA polymerase and a probe labelled with an environment sensitive, stable and fluorescent lanthanide chelate. In this assay, a non-competitive, exogenous internal standard is used. Both assays have a wide linear range (50-5 × 106 and 10-5 × 107 input PSA cDNA molecules for the end-point and real-time assays, respectively) and there is a strong correlation between the results obtained with the two assays (r = 1.0). Being somewhat faster to perform, the real-time format is better suited for assays that require high throughput.
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