HM Medical Clinic

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Enzymatic Immunoassay for the quantitative determination of free Prostata Specific Antigen (PSA) in human serum The standards are calibrated against NIBSC (WHO) Prostate cancer is the most frequent type of cancer found in the man and the second cause of death due to the cancer in man, Until recently, digital rectal examination was the most accepted Zero Standard/Diluent: diagnostic modality for early stage of prostate cancer but the ready-to-use reagent (10ml) measurement of serum PSA has become the most accepted test to indicate men who are at risk of having prostate cancer and who should be examined by other tests. Other biochemical ready-to-use reagent (0.50 ml) markers used in this context (PAP, total alkaline phosphatase, for concentration see label. carcinoembryonic antigen, etc,), are not as specific for prostate cancer. In 1979, Wang et al isolated a specific antigen for normal Peroxidase Conjugate: prostate tissue and called this protein PSA. As demonstrated by ready to use reagent (6 ml) Immunohistological studies, PSA is localized in the cytoplasm of prostate acinar cells, ductal epithelium and in the secretion on the ductal lumina, present in normal, benign hyperplastic and malignant prostate tissues as well metastatic prostate cancer ready to use reagent (6 ml) and in seminal plasma. An elevation of the serum concentration is reported in patients with both benign prostatic hypertrophy Prostate carcinoma, but rarely in healthy men and is absent in Concentrate – 25 ml (40 fold) normal women, The recent literature indicates that serum testing for PSA has become a very important tool to screen for prostate TMB – Substrate cancer and to monitor therapy of this disease, Serial ready-to-use reagent (12ml) measurement of PSA concentration in the serum is an important tool in monitoring patients with prostate cancer and determining the potential and actual effectiveness of surgery or other therapies, or may allow for earlier discovery of residual or recurrent carcinoma after radical prostatectomy or radiotherapy. Storage and Stability Priniciple of the Test Store kit at 2 - 8°C This assay is a solid phase enzyme-linked immunosorbent assay For expiry date see label of the test kit. (ELISA) based on the sandwich principle. The microtiter wells are coated with an antibody, directed towards an epitope of an The assay calibrators and controls are of human origin and have antigen molecule. An aliquot of patient serum is incubated in the been tested and confirmed negative for HIV, HBsAg and HCV by coated well with enzyme conjugated second antibody (E-Ab), FDA approved procedures. All standards, however, should be directed towards a different region of the antigen molecule. After treated as potential biohazards in use and for d isposal. The incubation the unbound E-Ab is washed off, The amount of assay reagents contain sodium azide or thimerosal which may bound E-Ab is proportional to the concentration of antigen in the be toxic if ingested. Sodium azide may react with copper and sample. After adding the substrate solution, the intensity of lead piping to form highly explosive salts. On disposal, flush with colour developed is proportional to the antigen concentration in large quantities of water. The stop solution contains H the sample. The measured Ods of the standards are used to comes into contact with skin, wash thorougly with water and seek construct a calibration curve against which the unknown samples medical attention. Since the H 2SO4 used to terminate the color reaction is corrosive, the instrumentation employed to dispense it should be thorougly cleaned after use. This kit is for in vitro diagnostic use only. Never pipette by mouth and avoid contact of reagents and specimens with skin and mucous membranes. If Each kit contains reagents sufficient for 96 determinations. contact occurs, wash with a germicidical soap and copious amounts of water. Do not smoke, eat or drink in areas where Microtiterplate: specimens or kit reagents are handled. Wear disposable latex 12 modules with 8 wells each (96 determinations) gloves when handling specimens and wash hands thoroughly afterwards. Microbial contamination specimens may give false 5 Free PSA-Standards: ready-to-use reagents (0.50 ml) at the following Material required but not provided: 12 ng PSA/mL; 6 ng PSA/mL; 3 ng PSA/mL; 1.5 ng PSA/mL; 0.75 ng PSA/mL • Precision micropipettes (volume: 25µl and 100µl) with • Distilled water • ELISA photometer with 450nm- and 630nm-filters • Timer with 60 min. range or higher • Microplate washer • Vortex or similar mixing tools • Container for the proper handling of waste and samples after Guideline for the preparation of samples Either fresh sera or plasma can be used. If not used immediately they can be stored at 2-8°C for 1 week: in case of longer storage, freeze at -20°C. Samples should be without microbial contaminations. In case, centrifuge at 2000g for 20 minutes or filtrate with a 0.2µ filter. Highly lipemic or hemolized samples can give uncorrect analytical results. Samples containing high titles of rheumatoid factor and anti-mouse heterophylous antibodies could give false positive results. It is recommended that internal controls be used in every assay. Control results should be within established ranges. Generally the concentration of PSA goes down, even if samples are stored at -20°C. Healthy men have Free PSA concentrations of 0.3 ng/mL or less. Normal Value and Cut-off Value: 1) Dilute 25mL Wash buffer in 975mL distilled Water 2) Prior to use bring all reagents to room temperature (18-25°C) 0.1 ± 0.25 ng Free PSA/mL 3) Pipette 25µl of e.g. standards, controls or samples into each well. Samples with an expected PSA value higher than 0.5 ng Free PSA/mL 12ng/ml should be diluted with the diluent. Note: The above values are only for users guidance. Every 4) Add 50 µl of Assay Reagent into each well and mix by laboratory should determine its own values. moving plate on the table (10 sec.) PERFORMANCE CHARACTERISTICS 5) Incubate 1 h at room temperature (18-25°C) 6) Add 50 µl of peroxidase conjugate into each well and mix by The limit of detection for this kit is 0.1 ng/mL moving plate on the table (10 sec.) 7) Incubate 1 h at room temperature (18-25°C) Intra- and inter-assay precisions were established by analysing three patient sera of different PSA concentrations. 8) Remove solution from the wells by aspirating or tapping the The results are shown in Tables 1 and 2. 9) For washing fill plate with diluted wash buffer and wait 10sec. Table 1) Intra-assay precision before removing the Wash Buffer; repeat wash 5 times (for a total of 6 times) 10) Pipette 100µl TMB-substrate solution into each well 11) Incubate 15min. at room temperature (18-25°C) 12) Add 100µl/well stopping solution (same order as substrate Table 2) Inter-assay precision 1) Calculate the mean absorbance for each duplicate. 2) Substract the absorbance value of the zero standard from the mean absorbance values of standards, control and samples. 3) Draw the standard curve on lin-lin or log-log graph paper by plotting absorbance values of standards against appropriate PSA concentrations. Spiked serum samples were prepared by adding aliquots of 4) Read off the PSA concentrations for the control and the samples with highly elevated free PSA to normal male serum samples. The recovery of the antigen was in between 91.8 – 116% and in average 102%. Worksheet and standard curve of typical assay: Not to be used for calculation of actual test results. Hook effect No hook effect has been noticed with samples up to 5000ng/mL. The BIOTINA total PSA ELISA was compared to a commercially available and CE marked two step Free PSA Elisa: Y = 0.8922x + 0,11 Do not expose to sunlight Specificity. The antibodies used in this kit are highly specific for Free PSA, with a relatively low cross-reactivity to other proteins and polypeptides, lipids or chemotherapeutic agents present in patient samples. Storage temperature Table 3) Specificity Read user instructions carefully TUV Product Service GmbH, Germany Rev. 10.11.2005BEF Elseyer Str. 59, 58119 Hagen, Germany Doxorubicin * HCl Diethylstilbestrol Fritsche, H,A, und R,J. Babalan; Analytical performance goals for measuring prostate-specific antigen: Clin, Chem, 39,1529-1529 (1993) Lange, p, et al,: The value of serum prostate-specific antigen determinations before and after radical prostatectomy, J, Urol, 141,873 (1989) Stamey, T, et al,; Prostate-specific antigen as serum marker for adenocarcinoma of the prostate, N, Engl, J, Med, 317,909-916 (1987) In-Vitro Diagnostic Kit


The impact of new social media on intercultural adaptation

University of Rhode Island The Impact of New Social Media on Intercultural Follow this and additional works at: Part of the nd the Recommended CitationSawyer, Rebecca, "The Impact of New Social Media on Intercultural Adaptation" (2011). Senior Honors Projects. Paper 242. This Article is brought to you for free and open access by the Honors Program at the University of Rhode Island at DigitalCommons@URI. It has beenaccepted for inclusion in Senior Honors Projects by an authorized administrator of DigitalCommons@URI. For more information, please contact.

Nonparametric bayes modelling of count processes

Nonparametric Bayes modelling of count processes © 2013 by Antonio Canale and David B. Dunson. Any opinions expressed here are those of the authorsand not those of the Collegio Carlo Alberto. Biometrika Advance Access published October 5, 2013 Biometrika (2013), pp. 1–16 Printed in Great Britain Nonparametric Bayes modelling of count processes