Development and evaluation of three real-time immuno-PCR assemblages for quantification of PSA

Lind, Kristina; Kubista, Mikael

doi:10.1016/j.jim.2005.06.015

Cited by 93 publications

(64 citation statements)

References 17 publications

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“…The imprecision of our assay is higher than that of conventional immunoassays. A similar imprecision has been reported for other immunoassays using PCR amplification (17,25,26 ). In spite of this, the assay for PSA-API is of potential clinical utility.…”

Section: Discussionsupporting

confidence: 72%

Proximity Ligation Measurement of the Complex between Prostate Specific Antigen and α1-Protease Inhibitor

et al. 2009

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Background: Prostate specific antigen (PSA)–α1-protease inhibitor complex (PSA-API) is a minor form of PSA in serum. It may be useful for prostate cancer (PCa) diagnosis, but its specific detection is hampered by nonspecific background. To avoid this, we developed an immunoassay for PSA-API based on proximity ligation. Methods: We used a monoclonal antibody (mAb) to total PSA (tPSA) to capture PSA, while using another anti-tPSA mAb together with an anti-API mAb as probes. We measured PSA-API by quantification of amplified DNA strands conjugated to the probes. We measured serum PSA-API in 84 controls and 55 men with PCa who had PSA concentrations of 4.0–10 μg/L. Results: The detection limit of the assay was 6.6 ng/L. The proportion of PSA-API to tPSA (%PSA-API) tended to be lower in men with PCa (2.8%) than without cancer (3.3%) but was not statistically significant (P = 0.363). When used alone, %PSA-API [area under the curve (AUC) 0.546] did not improve detection of PCa, whereas %fPSA (AUC 0.710) and the sum of %fPSA and %PSA-API (AUC 0.723) did. At 90% diagnostic sensitivity, the diagnostic specificity for cancer was not significantly better for %fPSA + %PSA-API than for %fPSA alone (36% vs 30%). Conclusions: Proximity ligation eliminated nonspecific background, enabling accurate measurement of PSA-API in serum specimens with moderately increased tPSA. The combined use of %PSA-API and %fPSA provided a modest improvement for PCa detection, but based on the current study cohort, it is uncertain whether the improvement has clinical utility. .

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Section: Discussionsupporting

confidence: 72%

Proximity Ligation Measurement of the Complex between Prostate Specific Antigen and α1-Protease Inhibitor

et al. 2009

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“…The concentration of extracellular IL-17 in cell-free BALF was analysed using immuno-qPCR [17]. The IL-17 immuno-qPCR was developed specifically for the present study in collaboration with a commercial biotechnology company (TATAA Biocenter, Gothenburg, Sweden).…”

Section: Analysis Of Soluble Balf Cytokinesmentioning

confidence: 99%

Interleukin-17-producing T-helper cells and related cytokines in human airways exposed to endotoxin

Glader¹,

Smith²,

Malmhäll³

et al. 2010

Eur Respir J

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Previous studies on mouse models have indicated that interleukin (IL)-17 and IL-17-producing T-helper (Th) cells are important for pulmonary host defence against Gram-negative bacteria. Human correlates to these findings have not yet been demonstrated. The aim of the present study was to determine whether or not IL-17-producing Th cells are present and whether IL-17 and other Th17-associated cytokines are involved in the immunological response to endotoxin in human airways.Segmental exposure to endotoxin and contralateral exposure to vehicle were performed in the lungs of healthy volunteers, with subsequent bronchoalveolar lavage 12 or 24 h after exposure to study local changes in cytokines and inflammatory cells.Endotoxin exposure increased concentrations of IL-17, IL-22 and their downstream effector molecules, human b-defensin-2 and IL-8/CXC chemokine ligand 8, in bronchoalveolar lavage fluid. Th cells with the capacity to produce IL-17 were found among the bronchoalveolar lavage cells, and expression of IL-17 mRNA correlated with expression of the transcription factor, retinoic-acidreceptor-related orphan receptor C variant 2. Moreover, endotoxin increased the numbers of neutrophils, macrophages and IL-17-producing T-cells, as well as the concentration of the Th17-regulating cytokines, IL-21 and IL-23.In conclusion, IL-17-producing Th cells are present, and IL-17, as well as other Th17-associated cytokines, is involved in the immunological response to endotoxin in human airways.

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“…1,2 Several methods for PSA detection are reported. [3][4][5][6][7][8][9][10][11][12] In men, PSA is expressed by prostate gland cells. In women, it is secreted in the periurethal gland 13 and in the breast.…”

Section: Introductionmentioning

confidence: 99%

Arraying prostate specific antigen PSA and Fab anti‐PSA using light‐assisted molecular immobilization technology

et al. 2010

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We here report for the first time the creation of prostate specific antigen (PSA) and Fab anti-PSA biosensor arrays using UV light-assisted molecular immobilization (LAMI), aiming at the detection and quantification of PSA, a cancer marker. The technology involves formation of free, reactive thiol groups upon UV excitation of protein aromatic residues located in spatial proximity of disulphide bridges, a conserved structural feature in both PSA and Fab molecules. The created thiol groups bind onto thiol reactive surfaces leading to oriented covalent protein immobilization. Protein activity was confirmed carrying out immunoassays: immobilized PSA was recognized by Fab anti-PSA in solution and immobilized Fab anti-PSA cross-reacted with PSA in solution. LAMI technology proved successful in immobilizing biomedically relevant molecules while preserving their activity, highlighting that insight into how light interacts with biomolecules may lead to new biophotonic technologies. Our work focused on the application of our new engineering principles to the design, analysis, construction, and manipulation of biological systems, and on the discovery and application of new engineering principles inspired by the properties of biological systems.

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Development and evaluation of three real-time immuno-PCR assemblages for quantification of PSA

Cited by 93 publications

References 17 publications

Proximity Ligation Measurement of the Complex between Prostate Specific Antigen and α1-Protease Inhibitor

Proximity Ligation Measurement of the Complex between Prostate Specific Antigen and α1-Protease Inhibitor

Interleukin-17-producing T-helper cells and related cytokines in human airways exposed to endotoxin

Arraying prostate specific antigen PSA and Fab anti‐PSA using light‐assisted molecular immobilization technology

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