scholarly journals Development of immunoradiometric assay for quantitative determination of free prostate-specific antigen

2005 ◽  
Vol 24 (2) ◽  
pp. 129-134 ◽  
Author(s):  
Miroslava Jankovic ◽  
Maja Kosanovic ◽  
Ljiljana Hajdukovic-Dragojlovic ◽  
Snezana Golubovic
Keyword(s):  
Quantitative Determination ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Laboratory Diagnostics ◽  
Immunoradiometric Assay ◽  
Serum Samples ◽  
Coefficients Of Variation ◽  
One Step ◽  
Prostate Diseases

In this study we reported the development and analytical validation of new assay for quantitative determination of free prostate-specific antigen, fPSA. It is formulated as one step, two-site "sandwich" immunoradiometric assay. Specificity of this assay was achieved by using epitope-1-reactive anti-fPSA antibody as tracer antibody. Assay was calibrated against first international standard 96/668, and its detection limit was determined as 0.08 mg/L. Intra- and inter-assay coefficients of variation were 3.42-7.53% and 7.04-8.33%, respectively. Measured concentrations of serially diluted serum samples were close to the calculated concentrations, indicating good linearity with recovery percentage ranging from 98.7-107.4%. Analytical performance characteristics of fPSA assay speaks in favor of its use as a reliable tool in laboratory diagnostics relating to prostate diseases.

scholarly journals Prostate Specific Antigen Detection Using Microgapped Electrode Array Immunosensor with Enzymatic Silver Deposition

2009 ◽  
Vol 55 (5) ◽  
pp. 964-971 ◽  
Author(s):  
Yong Huang ◽  
Tai-Hong Wang ◽  
Jian-Hui Jiang ◽  
Guo-Li Shen ◽  
Ru-Qin Yu
Keyword(s):  
Prostate Specific Antigen ◽  
Limit Of Detection ◽  
Specific Antigen ◽  
Electrical Conductance ◽  
Electrode Array ◽  
Serum Samples ◽  
Silver Deposition ◽  
Low Concentrations ◽  
Chemiluminescent Immunoassay ◽  
Prostate Diseases

Abstract Background: Analysis of trace proteins plays an essential role in the fields of biomedical research and clinical diagnosis. Development of methods for the detection of proteins at very low concentrations has historically been a challenge in immunochemistry. We have developed an electrical immunosensor for the detection of prostate specific antigen (PSA). Methods: The electrical immunosensor uses a microgapped interdigitated electrode array (MGIDEA) based on enzymatic silver deposition reaction. The deposition of silver was dispersed over the microgaps and allows the microgapped interdigitated electrodes to be electrically connected, resulting in an increase in electrical conductance of MGIDEA that is used to quantify the analyte concentration. We used this electrical immunosensor to measure PSA in human serum samples from patients with prostate diseases. Results: This electrical immunosensor exhibited a linear response with PSA concentrations over a 6-decade range from 1.0 pg/L to 1.0 μg/L, with detection limit of 0.9 pg/L. PSA concentrations using this immunosensor agreed within 10% of those obtained using a commercial chemiluminescent immunoassay. Conclusions: The MGIDEA method has characteristics (analyte specific, low background, low limit of detection) that provide potential for molecular detection in various biomedical areas.

scholarly journals Urinary prostate specific antigen: Is the clinical use likely?

2005 ◽  
Vol 52 (4) ◽  
pp. 69-74 ◽  
Author(s):  
T. Pejcic ◽  
J. Hadzi-Djokic ◽  
M. Acimovic ◽  
C. Topuzovic ◽  
B. Milkovic ◽  
...  
Keyword(s):  
Healthy Volunteers ◽  
Prostate Specific Antigen ◽  
Androgen Deprivation Therapy ◽  
Specific Antigen ◽  
Androgen Deprivation ◽  
Trus Biopsy ◽  
Significant Difference ◽  
Prostate Diseases

Prostate specific antigen (PSA) blood test represents the standard procedure in prostate cancer (CaP) diagnosis and follow-up. However, determination of PSA in the urine, where PSA is present in much higher concentrations than in the blood, still remains in the field of research. Objectives: To determine urinary concentrations of PSA (uPSA) in different groups of patients (pts.), and to estimate is it possible to differentiate benign and malignant prostate diseases and to follow-up the results of treatment. Methods: Between January 2001. and November 2003., urinary concentrations of PSA were determined at 142 pts. divided in seven groups: 1. young and healthy volunteers, 2. "BPH-24": pts. with benign prostatic hyperplasia (BPH) who collected the sample of 24- hour voided urine, 3. "BPH-I": pts. with BPH who collected the first portion of first urinary voiding, 4. "TRUS-CaP": pts. with CaP which gave the first portion of urine just prior to transrectal ultrasound guided prostate biopsy (TRUS- biopsy), 5. "TRUSnon- CaP": pts. who gave first portion of urine prior to TRUS-biopsy, but biopsy did not prove the presence of CaP, 6. "RRP": pts. who underwent radical retropubic prostatectomy (RRP), 7. "AAT": pts. who underwent androgen deprivation therapy. Results: Average uPSA value in the group of young and healthy volunteers, was 13.8+19.6 ng/ml, in "BPH-24": 38.0+ 44.4 ng/ml, in "BPH-I": 140.8+140.9 ng/ml, in "TRUSCaP": 234.8+277.7 ng/ml, in TRUS-non-CaP: 113.1 +148.5 ng/ml, and in the group "RRP": 4.4+4.7 ng/ml. There was no statistically significant difference of average uPSA values between "BPH-I" and "TRUSCaP" groups. The significant difference was found between the group of young volunteers and "BPH-I". In "TRUS-CaP" group, there was strong correlation between tumor size and aggressiveness and uPSA concentration. Finally, PSA and uPSA decline during androgen deprivation therapy, strongly correlated (up to r=0.95). Conclusions: Determination of uPSA cannot differentiate BPH and CaP. However, in the group of pts. with proven localized CaP, uPSA can provide additional information concerning T-staging. Moreover, simultaneous monitoring of PSA and uPSA response on hormonal therapy, can provide an early recognition of androgen-in different CaP (AIPCA) and hormone resistant CaP (HRPCA).

scholarly journals Measurement of the Complex between Prostate-specific Antigen and α1-Protease Inhibitor in Serum

1999 ◽  
Vol 45 (6) ◽  
pp. 814-821 ◽  
Author(s):  
Wan-Ming Zhang ◽  
Patrik Finne ◽  
Jari Leinonen ◽  
Satu Vesalainen ◽  
Stig Nordling ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Protease Inhibitor ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Serum Samples ◽  
Free Psa ◽  
Total Psa ◽  
Healthy Females

Abstract Background: Prostate-specific antigen (PSA) occurs in serum both free and in complex with protease inhibitors. The complex with α1-antichymotrypsin (ACT) is the major form in serum, and the proportion of PSA-ACT is higher in prostate cancer (PCa) than in benign prostatic hyperplasia (BPH). PSA also forms a complex with α1-protease inhibitor (API) in vitro, and the PSA-ACT complex has been detected in serum from patients with prostate cancer. The aim of the present study was to develop a quantitative method for the determination of PSA-API and to determine the serum concentrations in patients with PCa and BPH. Methods: The assay for PSA-API utilizes a monoclonal antibody to PSA as capture and a polyclonal antibody to API labeled with a Eu-chelate as a tracer. For calibrators, PSA-API formed in vitro was used. Serum samples were obtained before treatment from 82 patients with PCa, from 66 patients with BPH, and from 22 healthy females. Results: The concentrations of PSA-API are proportional to the concentrations of total PSA. PSA-API comprises 1.0–7.9% (median, 2.4%) of total immunoreactive PSA in PCa and 1.3–12.2% (median, 3.6%) in BPH patients with serum PSA concentrations >4 μg/L. In patients with 4–20 μg/L total PSA, the proportion of PSA-API serum is significantly higher in BPH (median, 4.1%) than in PCa (median, 3.2%; P = 0.02). Conclusions: The proportion of PSA-API in serum is lower in patients with PCa than in those with BPH. These results suggest that PSA-API is a potential adjunct to total and free PSA in the diagnosis of prostate cancer.

scholarly journals Dual-Label Immunoassay for Simultaneous Measurement of Prostate-specific Antigen (PSA)-α1-Antichymotrypsin Complex Together with Free or Total PSA

2003 ◽  
Vol 49 (1) ◽  
pp. 97-103 ◽  
Author(s):  
Lei Zhu ◽  
Jari Leinonen ◽  
Wan-Ming Zhang ◽  
Patrik Finne ◽  
Ulf-Håkan Stenman
Keyword(s):  
Detection Limit ◽  
Prostate Specific Antigen ◽  
Simultaneous Measurement ◽  
Solid Phase ◽  
Specific Antigen ◽  
Cathepsin G ◽  
Serum Samples ◽  
Total Psa ◽  
Dual Label

Abstract Background: A major portion of prostate-specific antigen exists in circulation as a complex with α1-antichymotrypsin (PSA-ACT), whereas a minor part is free (fPSA). The proportion of PSA-ACT is increased in prostate cancer (PCa), but immunologic determination of PSA-ACT is hampered by a background produced by nonspecific adsorption of ACT to the solid phase. To reduce the nonspecific interference, we produced an antibody specific for complexed ACT and developed immunofluorometric assays (IFMAs) for simultaneous measurement of fPSA + PSA-ACT (fPSA/PSA-ACT) and PSA-ACT + total PSA (tPSA, PSA-ACT/tPSA). Methods: Monoclonal antibodies (MAbs) were produced by immunization with PSA-ACT. The dual-label time-resolved IFMAs for fPSA/PSA-ACT and PSA-ACT/tPSA used a capture MAb to tPSA, an Eu3+-labeled MAb to fPSA or complexed ACT, and an Sm3+-labeled MAb to complexed ACT or to tPSA as tracer antibodies. The clinical utility was evaluated using serum samples from individuals with or without PCa with PSA concentrations of 2.0–20.0 μg/L. Results: One MAb (1D10) showed low cross-reactivity with free ACT and cathepsin G-ACT. A sandwich assay for PSA-ACT with 1D10 as tracer had a detection limit of 0.05 μg/L, and with this assay, PSA-ACT was undetectable in female sera. The detection limit for fPSA was 0.004 μg/L. Determinations of the ratio of fPSA to PSA-ACT and the proportions of fPSA/tPSA and PSA-ACT/tPSA provided the same clinical specificity for PCa and provided significantly better clinical specificity than did tPSA. Conclusions: Background problems observed in earlier PSA-ACT assays are eliminated by the use of a MAb specific for complexed ACT as a tracer. The same clinical validity can be obtained by determination of fPSA or PSA-ACT together or in combination with tPSA.

scholarly journals A one-step method for quantitative determination of uracil in DNA by real-time PCR

2010 ◽  
Vol 38 (21) ◽  
pp. e196-e196 ◽  
Author(s):  
András Horváth ◽  
Beáta G. Vértessy
Keyword(s):  
Quantitative Determination ◽  
Real Time ◽  
Real Time Pcr ◽  
Step Method ◽  
One Step

Measurement of total and free prostate specific antigen (PSA) in human serum samples using an ultra-microanalytical system

2021 ◽  
pp. 114470
Author(s):  
Juliette M. Cazanave Mora ◽  
Ruben del Valle García ◽  
Lilian Pérez López ◽  
Dunia C. Bequer Ariza ◽  
Orlando Zulueta Rodríguez ◽  
...  
Keyword(s):  
Human Serum ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Serum Samples ◽  
Human Serum Samples
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