Sirolimus西罗莫司检测试剂盒电化学发光法说明书

Elecsys Anti-HBs IIREFSYSTEM********************** 300cobas e 801EnglishSystem information Short name ACN (application code number) AHBS 2 10138 Immunoassay for the in vitro quantitative determination of human antibodies to the hepatitis B surface antigen (HBsAg) in human serum and plasma.Anti-HBs assays are used within the scope of hepatitis B vaccination to check the necessity and success of vaccination. In addition, anti-HBs assays are used to monitor the course of disease following acute hepatitis B infection. This test is not intended for diagnosis.The e lectro c hemi l uminescence i mmuno a ssay “ECLIA” is intended for use on the cobas e 801 immunoassay analyzer.Note: Please note that the catalogue number appearing on the package insert retains only the first 8 digits of the licensed 11-digit Catalogue Number: 07026854190 for the Elecsys Anti-HBs II assay. The last 3 digits -190 have been replaced by -119 for logistic purposes. SummaryAnti-HBs is a specific (generally IgG) antibody that is directed against the hepatitis B surface antigen (HBsAg).1,2 Anti ‑HBs can be detected several weeks after the disappearance of hepatitis B surface antigen.3,4 Anti ‑HBs can be formed following a hepatitis B infection or after hepatitis Bvaccination.3,4 Antibodies are formed against the HBsAg determinant a, which is common to all subtypes, and against subtype-specific determinants.1,5,6Anti ‑HBs assays are used within the scope of hepatitis B vaccination to check the necessity and success of vaccination.2,4,7 In addition, anti ‑HBs assays are used to monitor the course of disease following acute hepatitis B infection.3The Elecsys Anti ‑HBs II assay uses a mixture of purified antigens fromhuman serum (HBsAg subtype ad), and recombinant HBsAg subtype ay from CHO (Chinese Hamster Ovary) cells. Test principleSandwich principle. Total duration of assay: 18 minutes.▪ 1st incubation: Anti ‑HBs in the sample (24 μL), biotinylated HBsAg(ad/ay), and HBsAg (ad/ay) labeled with a ruthenium complex a) react to form a sandwich complex.▪ 2nd incubation: After addition of streptavidin-coated microparticles, thecomplex becomes bound to the solid phase via interaction of biotin and streptavidin.▪ The reaction mixture is aspirated into the measuring cell where themicroparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell II M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.▪ Results are determined via a calibration curve which is instrument-specifically generated by 2‑point calibration and a master curve provided via the cobas link.a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy)32+)Reagents – working solutionsThe cobas e pack (M, R1, R2) is labeled as AHBS 2. M Streptavidin-coated microparticles, 1 bottle, 13.2 mL:Streptavidin-coated microparticles 0.72 mg/mL; preservative. R1 HBsAg~biotin, 1 bottle, 16.7 mL:Biotinylated HBsAg (ad/ay) human/recombinant, > 0.5 mg/L; MES b) buffer 85 mmol/L, pH 6.5; preservative. R2 HBsAg~Ru(bpy)32+, 1 bottle, 15.8 mL:HBsAg (ad/ay) human/recombinant, labeled with ruthenium complex > 0.3 mg/L; MES buffer 85 mmol/L, pH 6.5; preservative.b) MES = 2-morpholino-ethane sulfonic acidAHBS 2 Cal1 Calibrator 1, 1 bottle of 1.3 mL:Anti ‑HBs (human) in human serum; preservative.AHBS 2 Cal2 Calibrator 2, 1 bottle of 1.3 mL:Anti ‑HBs (human) in human serum; preservative.Precautions and warnings For in vitro diagnostic use.Exercise the normal precautions required for handling all laboratory reagents. Disposal of all waste material should be in accordance with local guidelines. Safety data sheet available for professional user on request.This kit contains components classified as follows in accordance with the Regulation (EC) No. 1272/2008:n ‑Octyl ‑N,N ‑dimethyl ‑3‑ammonio ‑1‑propanesulfonateEUH 208 May produce an allergic reaction.Product safety labeling primarily follows EU GHS guidance. All human material should be considered potentially infectious.The calibrators (AHBS 2 Cal1 and AHBS 2 Cal2) have been preparedexclusively from the blood of donors tested individually and shown to be free from HBsAg and antibodies to HCV and HIV. The testing methods applied were FDA ‑approved or cleared in compliance with the European Directive 98/79/EC, Annex II, List A.The HBsAg starting material used was inactivated prior to labeling with biotin or ruthenium by heating to 60 °C for 15 hours. In addition, any virus particles remaining were removed by ultracentrifugation.However, as no inactivation or testing method can rule out the potential risk of infection with absolute certainty, the material should be handled with the same level of care as a patient specimen. In the event of exposure, the directives of the responsible health authorities should be followed.8,9 Avoid foam formation in all reagents and sample types (specimens, calibrators and controls). Reagent handlingThe reagents (M, R1, R2) in the kit are ready-for-use and are supplied in cobas e packs. CalibratorsThe calibrators are supplied ready ‑for ‑use in bottles compatible with the system.Unless the entire volume is necessary for calibration on the analyzer, transfer aliquots of the ready ‑for ‑use calibrators into empty snap ‑cap bottles (CalSet Vials). Attach the supplied labels to these additional bottles. Store the aliquots at 2‑8 °C for later use.Perform only one calibration procedure per aliquot.All information required for correct operation is available via the cobas link. Storage and stability Store at 2‑8 °C. Do not freeze.Store the cobas e pack upright in order to ensure complete availability of the microparticles during automatic mixing prior to use. Stability of the cobas e pack: unopened at 2‑8 °Cup to the stated expiration date on the cobas e 801 analyzer 16 weeksStability of the calibrators: unopened at 2‑8 °C up to the stated expiration date after opening at 2‑8 °C 16 weeks on the cobas e 801 analyzer at 20‑25 °Cuse only onceadhering to the snap ‑cap.Specimen collection and preparationOnly the specimens listed below were tested and found acceptable.Serum collected using standard sampling tubes or tubes containing separating gel.K2‑EDTA and K3‑EDTA plasma.Criterion: Slope 1.00 ± 0.15 + intercept 0 ± 2 IU/L + bias at 10 IU/L: ≤ 30 %. Stable for 3 days at 20‑25 °C, 6 days at 2‑8 °C, 3 months at ‑20 °C(± 5 °C). The samples may be frozen 5 times.For plasma treated with lithium heparin, lithium heparin with gel or sodium heparin, the values found were on average up to 20 % lower than those obtained in serum. For plasma treated with sodium citrate, the values found were on average up to 30 % lower than those obtained with serum.The sample types listed were tested with a selection of sample collection tubes or systems that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.Centrifuge samples containing precipitates and thawed samples before performing the assay.Do not use heat‑inactivated samples.Do not use samples and controls stabilized with azide.Ensure the samples and calibrators are at 20‑25 °C prior to measurement. Due to possible evaporation effects, samples and calibrators on the analyzers should be analyzed/measured within 2 hours.The performance of the Elecsys Anti‑HBs II assay has not been established with cadaveric samples or body fluids other than serum and plasma. Materials providedSee “Reagents –working solutions” section for reagents.▪ 2 x 6 bottle labelsMaterials required (but not provided)▪REF 11876317122, PreciControl Anti‑HBs, 16 x 1.3 mL▪REF 11776576322, CalSet Vials, 2 x 56 empty snap-cap bottles▪REF***********,DiluentUniversal,45.2mLsamplediluent▪▪cobas e 801 analyzerAccessories for the cobas e 801 analyzer:▪REF***********,ProCellIIM,2x2Lsystemsolution▪REF 04880293190, CleanCell M, 2 x 2 L measuring cell cleaning solution ▪REF***********,ReservoirCups,8cupstosupplyProCellIIMand CleanCell M▪REF***********,PreCleanIIM,2x2Lwashsolution▪REF***********,AssayTip/AssayCuptray,6magazinesx6magazine stacks x 105 assay tips and 105 assay cups, 3 wasteliners▪REF***********,LiquidFlowCleaningCup,2adaptorcupstosupply ISE Cleaning Solution/Elecsys SysClean for Liquid Flow CleaningDetection Unit▪REF***********,PreWashLiquidFlowCleaningCup,1adaptorcupto supply ISE Cleaning Solution/Elecsys SysClean for Liquid Flow Cleaning PreWash Unit▪REF 11298500316, ISE Cleaning Solution/Elecsys SysClean,5 x 100 mL system cleaning solutionAssayFor optimum performance of the assay follow the directions given in this document for the analyzer concerned. Refer to the appropriate operator’s manual for analyzer‑specific assay instructions.Resuspension of the microparticles takes place automatically prior to use. Place the cooled (stored at 2‑8 °C) cobas e pack on the reagent manager. Avoid foam formation. The system automatically regulates the temperature of the reagents and the opening/closing of the cobas e pack. Calibrators:Place the calibrators in the sample zone.Read in all the information necessary for calibrating the assay.CalibrationTraceability: This method has been standardized against the 1st WHO Reference Standard 1977.The predefined master curve is adapted to the analyzer using AHBS 2 Cal1 and AHBS 2 Cal2.Calibration frequency: Calibration must be performed once per reagent lot using AHBS 2 Cal1, AHBS 2 Cal2 and fresh reagent (i.e. not more than24 hours since the reagent kit was registered on the analyzer).Renewed calibration is recommended as follows:▪after 12 weeks when using the same reagent lot▪after 28 days when using the same cobas e pack on the analyzer▪as required: e.g. quality control findings with PreciControl Anti‑HBs outside the defined limitsQuality controlFor quality control, use PreciControl Anti‑HBs.Controls for the various concentration ranges should be run individually at least once every 24 hours when the test is in use, once per cobas e pack, and following each calibration.The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.If necessary, repeat the measurement of the samples concerned.Follow the applicable government regulations and local guidelines for quality control.CalculationThe analyzer automatically calculates the analyte concentration of each sample in IU/L.Interpretation of the resultsNumeric result Result message Interpretation< 10 IU/L Non-reactive Negative for anti-HBs≥ 10 IU/L Reactive Positive for anti-HBsvary depending on the testing procedure used. Results obtained from a single sample using tests from different manufacturers can therefore differ by up to a factor of 4 (or even a factor of 10 in rare cases). If there is a change in the assay procedure used during the monitoring of vaccination protection, then the anti‑HBs values obtained upon changing over to the new method must be confirmed by parallel measurements by both methods. Vaccination strategies in certain risk groups are based on the measured anti‑HBs concentration. Respective recommendations are given by national or regional guidelines. Limitations - interferenceThe effect of the following endogenous substances and pharmaceutical compounds on assay performance was tested. Interferences were tested up to the listed concentrations and no impact on results was observed. Endogenous substancesCompound Concentration testedBilirubin ≤ 513 μmol/L or ≤ 30 mg/dL Hemoglobin ≤ 0.621 mmol/L or ≤ 1000 mg/dL Intralipid ≤ 1500 mg/dLBiotin ≤ 41 nmol/L or ≤ 10 ng/mL Rheumatoid factors ≤ 1200 IU/mLAlbumin ≤ 7.0 g/dLIgG ≤ 7.0 g/dLIgA ≤ 1.6 g/dLIgM ≤ 1.0 g/dL2 / 42017-09, V 1.0 Can EnglishCriterion: Recovery for samples from Limit of Detection to 10 IU/L:≤ ± 2 IU/L, and samples > 10 IU/L: ≤ ± 20 % of initial value.Samples should not be taken from patients receiving therapy with high biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin administration.Pharmaceutical substancesIn vitro tests were performed on 16 commonly used pharmaceuticals. No interference with the assay was found.In addition, the following special drugs used in hepatitis B therapy were tested. No interference with the assay was found.Special drugsDrug Concentration testedmg/LPeginterferon alfa‑2a ≤ 0.18Peginterferon alfa‑2b ≤ 1.6Lamivudine ≤ 300Adefovir ≤ 10Entecavir ≤ 10Tenofovir ≤ 600Telbivudine ≤ 245Due to high-dose hook effect c), results from anti‑HBs concentrations of> 200000 IU/L may be found below the upper limit of the measuring range of 1000 IU/L. In rare cases, a high-dose hook effect from anti HBs concentrations of < 20000 IU/L cannot be excluded. Therefore in case of any unexpected low result the sample should be diluted 1:100 (refer to chapter “Dilution”) and tested again.In rare cases, interference due to extremely high titers of antibodies to streptavidin and ruthenium can occur. The test contains additives which minimize these effects.c) High-dose hook effect: A sample with a true concentration clearly above the measuring range, but found within the measuring range.Limits and rangesMeasuring range2‑1000 IU/L (defined by the Limit of Detection and the maximum of the master curve). Values below the Limit of Detection are reported as< 2 IU/L.Values above the measuring range are reported as > 1000 IU/L (or up to 100000 IU/L for 100‑fold diluted samples).DilutionSamples with anti‑HBs concentrations above the measuring range can be diluted with Diluent Universal. The recommended dilution is 1:100 (either automatically by the analyzer or manually). The concentration of the diluted sample must be > 10 IU/L.After manual dilution, multiply the result by the dilution factor.After dilution by the analyzer, the software automatically takes the dilution into account when calculating the sample concentration.Manual dilution can also be made with negative human serum.Note: Antibodies to HBsAg are heterogeneous. In some isolated cases, this may lead to non-linear dilution behavior.Specific performance dataRepresentative performance data on the analyzer is given below. Results obtained in individual laboratories may differ.PrecisionPrecision was determined using Elecsys reagents, samples and controls in a protocol (EP05‑A3) of the CLSI (Clinical and Laboratory Standards Institute): 2 runs per day in duplicate each for 21 days (n = 84). The following results were obtained:cobas e 801 analyzerRepeatability d)Intermediateprecision e)Sample MeanIU/LSDIU/LCV%SDIU/LCV% Human serum 1 4.33 0.224 5.2 0.272 6.3 Human serum 2 12.0 0.237 2.0 0.277 2.3 Human serum 3 475 6.81 1.4 7.55 1.6 PC f) Anti-HBs 1 < 2.00 - - - -PC Anti-HBs 2 83.8 1.08 1.3 1.28 1.5d) Repeatability = within-run precisione) Intermediate precision = between-run precisionf) PC = PreciControlAnalytical specificityNo cross-reactions with HAV, HCV, HEV, CMV, EBV, HIV, Rubella, Toxoplasma gondii, Treponema pallidum, rheumatoid arthritis, autoimmune response or alcoholic liver disease were observed.Measurements were performed on each of the pathogens listed above using ≥ 8 serum or plasma samples which were positive for antibodies to the above-mentioned pathogens.Relative sensitivityPerformance of the Elecsys Anti‑HBs II assay has been assessed by testing a total of 669 samples at two different study sites. 296 samples from vaccinated persons and 373 samples from patients recovered from a hepatitis B infection have been measured with the Elecsys Anti‑HBs II assay and another commercially available fully automated anti‑HBs assay. Discrepant samples were tested with additional anti‑HBs assays to achieve a consensus.Characterization ofsamplesN ElecsysAnti‑HBs IIreactiveAnti‑HBscomparisontest reactiveSensitivity%Anti-HBs positive:vaccinees 296 296 296 100Anti-HBs positive:recovered from ahepatitis B infection373 373 373 100 Total 669 669 669 100 Relative specificityPerformance of the Elecsys Anti‑HBs II assay has been assessed by testing 2673 samples from blood donors negative for anti‑HBs at two different study sites and 1623 anti‑HBs negative samples from laboratory routine at three different study sites. Discrepant samples were tested with additional anti‑HBs assays to achieve a consensus.Characterization of samples N ElecsysAnti‑HBs IIfalsepositiveSpecificity%Anti-HBs negative: blood donors 2673 6 99.78 Anti-HBs negative: routinesamples1623 9 99.45 References1Seeger C, Zoulim F, Mason WS. Hepadnaviruses. In: Field’s Virology, Knipe DM, Howley RM (eds), 2007 5th edition, Lippincott Williams andWilkins, Philadelphia, USA. Chapter 76, pp2977-3029.2WHO. Hepatitis B vaccines. Wkly Epidemiol Rec 2009;84:405-420.3Liaw YF, Chu CM. Hepatitis B virus infection. Lancet2009;373:582-592.4Caspari G, Gerlich WH. The serologic markers of hepatitis B virus infection – proper selection and standardized interpretation. Clin Lab2007;53:335-343.5Kramvis A, Kew M, François G. Hepatitis B virus genotypes. Vaccine 2005;23:2409-2423.6Michel ML, Tiollais P. Hepatitis B vaccines: protective efficacy and therapeutic potential. Pathol Biol 2010;58:288-295.7Elgouhari HM, Abu-Rajab Tamimi TI, Carey WD. Hepatitis B virus infection: understanding its epidemiology, course, and diagnosis. Cleve Clin J Med 2008;75:881-889.8Occupational Safety and Health Standards: Bloodborne pathogens. (29 CFR Part 1910.1030). Fed. Register.9Directive 2000/54/EC of the European Parliament and Council of18 September 2000 on the protection of workers from risks related toexposure to biological agents at workFor further information, please refer to the appropriate operator’s manual for the analyzer concerned, the respective application sheets, the product information and the Method Sheets of all necessary components (if available in your country).A point (period/stop) is always used in this Method Sheet as the decimal separator to mark the border between the integral and the fractional parts of a decimal numeral. Separators for thousands are not used.SymbolsRoche Diagnostics uses the following symbols and signs in addition to those listed in the ISO 15223‑1 standard:CONTENT Contents of kitSYSTEM Analyzers/Instruments on which reagents can be used REAGENT ReagentCALIBRATOR CalibratorVolume after reconstitution or mixingGTIN Global Trade Item NumberCOBAS, COBAS E, ELECSYS and PRECICONTROL are trademarks of Roche. INTRALIPID is a trademark of Fresenius Kabi AB.All other product names and trademarks are the property of their respective owners. Additions, deletions or changes are indicated by a change bar in the margin.© 2016, Roche DiagnosticsRoche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim。

收稿日期 :2010－08－17作者单位 :1. 大连医科大学附属第二医院药剂科 , 大连 116027; 2. 大连医科大学药学院 , 大连 116044*通讯作者西罗莫司血药浓度测定方法的建立及在肝移植患者中的应用张宁 1, 吕慧怡 1*, 邓卅 2, 范青1[摘要 ]目的建立高效液相色谱法测定西罗莫司血药浓度 , 测定该药在肝移植患者中的药代动力学参数。

方法以沉淀 -萃取法处理全血样品 , 采用Hypersil ODS C 18(250mm ˑ 4. 6mm , 5μm 柱 , 柱温为50ħ , 乙腈 -甲醇 -水 (7. 5ʒ 62. 5ʒ 30 为流动相 , 流速 1. 2mL /min , 以 32-去甲基雷帕霉素为内标 , 在 276nm 波长处 , 检测西罗莫司血药浓度 , 并应用此方法测定 5例肝移植患者的西罗莫司血药浓度。

结果全血中西罗莫司浓度在 2. 0 50. 0ng /mL 范围内线性良好 (r =0. 9997 , 平均相对回收率为 99. 1%(n =5 。

5例肝移植患者的 C max 为 (12. 66ʃ4. 1 ng /mL , t 1/2为 (19. 2ʃ 11. 2 h , t max 为 (1. 8ʃ 0. 3 h 。

结论本方法快速、简便、准确 , 灵敏度高 , 重现性好 , 可用于肝移植患者的西罗莫司临床药物监测。

[关键词 ]西罗莫司 ; 高效液相色谱法 ; 血药浓度 ; 肝移植Determination of sirolimus in human whole blood by HPLC and the exploratory in liver transplantation ZHANG Ning 1, L Hui-yi 1*, DENG Sa 2, FAN Qing 1(1. Department of Pharmacy ,The Second Affiliated Hospital of Dalian Medical University , Dalian 116027, China ; 2. Department of Pharmacology , Dalian Medical University , Dalian 116044, China[Abstract ]Objective To establish an HPLC method to determine the concentration of sirolimus in wholeblood．MethodsThe whole-blood samples were pretreated by precipitation-extraction method , and sirolimus was de-termined by HPLC-UV．Hypersil ODS C 18analysis column (250mm ˑ 4. 6mm ,5μm was used , and the temperature was set at 50ħ ．The mobile phase was a mixture of acetonitrile-methanol-water (7. 5ʒ 62. 5ʒ 30 with a flow rate of 1. 2mL /min．The detection wavelength was set at 276nm．ResultsThe calibration curve was linear within the range 2. 050ng /mL (r =0. 9997 . The mean relative recovery was 99. 1%, and the mean absolute recovery was 82. 2%(n =5 ．The intra-day and the inter-day relative standard deviation did not exceed 3. 3%and 6. 3%respectively．In the five transplantationreceptors , C max was (12. 66ʃ 4. 1 ng /mL , t 1/2was (19. 2ʃ 11. 2 h , t max was (1. 8ʃ 0.3 h．Conclusion This method is simple , rapid , sensitive and suitable for therapeutic drug monitoring of sirolimus．Key words :Sirolimus ; HPLC ; Plasma drug concentration ; Liver transplantation西罗莫司 (Sirolimus , SRL , 商品名 Rapamune , 雷帕霉素 , 是一种大环内酯类免疫抑制剂 , 具有抗增殖、抗肿瘤及无肾毒性等特性 , 近年来逐渐应用于肝移植患者 , 使临床获得了防治肝移植术后肾功能不全、慢性排斥反应、肿瘤复发和提高移植物长期存活的新途径 [1]。

化学发光法 EMSA 检测试剂盒Chemiluminescent EMSA Detection Kit货号：KGS133 保存温度：-20℃ For Research Use Only说明书修订日期：2015.12.01一、产品简介 本产品是一种通过 Streptavidin-HRP 及 ECL 检测试剂来实现化学发光检测 Biotin 标记的 EMSA 探针的检测试剂盒。

同时本试剂盒也提供 EMSA 检测所需的结合缓冲液和上样缓冲液，及一些关键的相关试剂， 可以实现非同位素的 EMSA 检测。

本试剂盒采用了高质量的 Streptavidin-HRP Conjugate，HRP 和 Streptavidin 共价交联的比例大于 3， 这样比采用 Streptavidin 和 Biotin-HRP conjugate 两种试剂进行检测要更方便，并且灵敏度更高。

本试剂盒采用了非特异性结合比 avidin 更低的 strepatavidin，使检测结果背景更低灵敏度更高。

EMSA/Gel-Shift 结合缓冲液(5X)中含有 poly(dI-dC)等有效成分。

其中 poly(dI-dC)的浓度经过优化， 可以很好的消除蛋白和标记探针间的非特异性结合，同时又不会减弱目的转录因子和标记探针间的结合。

本试剂盒可以用于 20 个蛋白和探针的结合反应，并足够检测至少 2 块有生物素标记 EMSA 探针的膜。

二、组份列表产品组份规格（50T）EMSA/Gel-Shift 结合缓冲液(5X)100µlEMSA/Gel-Shift 上样缓冲液(无色，10X)100µlEMSA/Gel-Shift 上样缓冲液(蓝色，10X)100µlECL Reagent A28mlECL Reagent B28mlStreptavidin-HRP Conjugate50µl封闭液190ml洗涤液(5X)125ml检测平衡液125ml注：如果长期不用，整个试剂盒可-20℃保存，-20℃可以保存更长时间。

西尼罗河病毒检测试剂盒【用途】美国FOCUS西尼罗河（West Nile）病毒IGM ELISA，利用了西尼罗重组抗原检测人体血清或血浆中的西尼罗抗体。

本实验仅用于辅助诊断人感染了西尼罗病毒，不用于筛查血液或血液成分。

仅供专业人员体外诊断使用。

【代理商】广州健仑生物科技有限公司【概述】感染了西尼罗病毒会导致包括脑炎等一系列症状的疾病，西尼罗病毒在世界广泛传播，已在50多个国家检测出该病毒。

本试验经CDC提供的试剂检验与验证。

本试验使用了一种称为WNRA的重组抗原，它可以作为西尼罗病毒感染的快速血清学指标，WNRA蛋白是一种重组抗原，它是由含有西尼罗病毒两个抗原的序列多肽构成。

【实验原理】检测西尼罗病毒IgG/IgM ELISA 是基于两步夹心法原理制造的。

【提供材料】1.微孔板（96孔 12x8）：即用每孔均包被结合西尼罗重组抗原的单抗。

2－8℃下保存至保质期。

注意：WNRA和NCA已包被于微孔板。

2.IGM样品稀释液：1支 25ml 即用 2-8℃下保存至使用。

3.IGM阴性质控：1支 30ul 2-8℃下保存至使用。

要长时间储存，将其分装于小瓶里，在-70℃下储存。

4.IGM阳性质控：1支 30ul 2-8℃下保存至使用。

要长时间储存，将其分装于小瓶里，在-70℃下储存。

5.洗涤液（10X）：1瓶 120ml 2－8℃下保存至使用。

6.EnWash：1瓶 20ml 2－8℃下保存至使用。

7.酶联物：一瓶6ml 即用 2－8℃下保存至使用。

8.TMB底物液：1支9ml 即用 2－8℃下保存至使用。

9.终止液；1支6ml 即用2－8℃下保存至使用。

试剂应避免反复冻融。

【操作步骤】在使用前将所有试剂和样品平衡至室温，并轻轻倒转使其充分混匀。

实验准备：将10X的浓缩洗涤液稀释,将120ml的浓缩洗涤液加上1080ml的蒸馏水，摇匀，至无任何沉淀，稀释好的洗涤液最多可在室温下放置两个星期。

准备实验所需的板条，剩余的板条应尽快放入袋子里重新密封，在2－8℃下储存至保质期。

化学发光法说明书化学发光法说明书一、引言化学发光法是一种常用的分析技术，它通过特定的化学反应产生发光现象，并利用测量发光强度来确定待测物质的浓度。

本说明书将介绍化学发光法的原理、实验步骤、仪器设备及应用领域。

二、原理化学发光法的原理基于化学反应产生发光的特性。

当特定的化学物质与待测物质发生反应时，会产生激发态的化学物种。

随后，激发态的化学物种会通过辐射或非辐射的方式回到基态，释放出能量并产生光。

通过测量发光强度，可以间接推测出待测物质的浓度。

三、实验步骤1. 准备样品：将待测物质制备成合适的浓度，并与特定的试剂混合。

2. 反应：在适当的条件下，使待测物质与试剂发生化学反应，产生发光。

3. 测量：利用发光仪器测量发光强度，并记录下来。

4. 构建标准曲线：根据已知浓度的标准样品的发光强度，绘制标准曲线。

5. 计算待测物质浓度：通过待测样品的发光强度，在标准曲线上找到对应的浓度值，并计算待测物质的浓度。

四、仪器设备化学发光法的实验需要使用特定的仪器设备，主要包括：1. 发光仪：用于测量样品的发光强度。

2. 标准样品：已知浓度的样品，用于构建标准曲线。

3. 试剂：与待测物质发生反应，产生发光的化学试剂。

4. 试管或微孔板：用于混合样品和试剂，进行反应。

五、应用领域化学发光法广泛应用于许多领域，包括：1. 生物医学研究：用于检测生物标志物、药物浓度等。

2. 环境监测：用于测定水中重金属、有机物等的浓度。

3. 食品安全：用于检测食品中的农药残留、添加剂等。

4. 公共安全：用于检测爆炸物、毒品等危险物质。

六、实验注意事项1. 实验操作要规范，遵守实验室安全规定。

2. 样品和试剂要保持干净和干燥，避免受到外界污染。

3. 仪器设备要正确校准，确保测量结果准确可靠。

4. 实验过程中要注意控制反应条件，如温度、pH值等。

5. 标准曲线的制备要精确可靠，避免误差产生。

七、总结化学发光法作为一种敏感、快速和准确的分析方法，在科学研究和实际应用中具有重要的地位。

西罗莫司片（雷帕鸣）中文说明书警示语：具有免疫抑制作用，不推荐用于肝或肺移植患者•由于免疫抑制作用，本品可增加感染机会也可能引发淋巴瘤及其他恶性肿瘤。

由于免疫抑制作用，本品可增加感染机会也可能引发淋巴瘤。

有免疫治疗和管理肾移植经验的医师方可使用本品。

使用本品的病人，应在具有一定资质条件的医疗机构内接受管理。

负责维持治疗的医师，应该不断完善病人的随访信息。

（见【注意事项】）•西罗莫司作为免疫抑制剂用于肝移植或肺移植患者的安全性和疗效尚未明确，因此，不推荐在此类患者中使用。

（见【注意事项】）•肝移植一死亡率增加、移植物失去功能及肝动脉血栓形成（HAT）在一项对新接受肝移植的患者进行的试验中，发现西罗莫司与他克莫司联合使用与死亡率和移植物失去功能增加相关。

这些患者中许多在死亡时或临近死亡时有感染的迹象。

在该试验及另一项对新接受肝移植患者进行的试验中，西罗莫司与环抱素或他克莫司联合使用与HAT发生率升高相关，大部份HAT发生于移植后30天内，并且大多导致了移植物失去功能或死亡。

（见【注意事项】）・肺移植一气管吻合处开裂新肺移植患者接受包括西罗莫司在内的免疫抑制治疗，有气管吻合处开裂的病例报道，大部分为致命性。

（见【注意事项】）【通用名称】西罗莫司片【商品名称】雷帕鸣【英文名称】Siro1.imusTab1.ets【汉语拼音】Xi1.uomosiPian【成份】本品的主要成份为西罗莫司。

化学名称：（3S,6R,7E,9R z10R,12R,14S z15E,17E z19E,21S,23S,26R,27R,34aS）-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a・十六氢∙9,27∙二羟基∙3-［（1.R）∙2-［（1.S,3R,4R）-4.羟基3甲氧环己基卜1•甲基乙基卜10,21・二甲氧・6,8,12,14,20,26・六甲基・23,27•环氧・3H・口比咤并［2,IyH4］氧杂氮杂三H^一环烯・1,5,11,28,29（4H,6H,31H）∙戊酮化学结构式：分子式：C SI H79NO I3分子量：914.2本品辅料为：乳糖一水合物，聚乙二醇8000粉末，硬脂酸镁，滑石粉，聚乙二醇20000,单油酸甘油酯（60%）,药用釉料（虫胶），无水硫酸钙，微晶纤维素，蔗糖，二氧化钛，泊洛沙姆188,聚维酮（K29/32）,维生素E（&・八生育酚），巴西棕檎蜡和其他成份。

(19)中华人民共和国国家知识产权局(12)发明专利申请(10)申请公布号 (43)申请公布日 (21)申请号 201711090619.6(22)申请日 2017.11.08(71)申请人 北京化工大学地址 100029 北京市朝阳区北三环东路15号(72)发明人 乐园　沈煜栋　陈鹏　林谡轩　(74)专利代理机构 北京志霖律师事务所 11575代理人 张文祎(51)Int.Cl.A61K 9/20(2006.01)A61K 47/26(2006.01)A61K 47/38(2006.01)A61K 47/32(2006.01)A61K 47/36(2006.01)A61K 47/20(2006.01)A61K 31/436(2006.01)A61P 37/06(2006.01)(54)发明名称一种西罗莫司纳米片剂及其制备方法(57)摘要本发明公开了一种西罗莫司纳米片剂包括如下重量份数的原料：西罗莫司纳米药物组合物2-15份，赋形剂80-160份，填充剂80-120份，崩解剂30-50份，粘合剂30-60份，润滑剂5-15份；本发明还公开了西罗莫司纳米片剂的制备方法。

本发明制备得到的西罗莫司纳米片剂，溶出度较进口片剂提高约15-30％，生物利用度更高，稳定性更好；便于包装、储存、运输及携带；便于吞服，服用方便，吸收性好，减少服用过程中药物损失。

本发明制备工艺过程简单，对设备无特殊要求，容易实现，能耗少，效率高，成本低，且非常容易放大，达到工业化生产的发明目标。

权利要求书1页 说明书8页 附图3页CN 107811979 A 2018.03.20C N 107811979A1.一种西罗莫司纳米片剂，其特征在于，包括如下重量份数的原料：西罗莫司纳米药物组合物 2-15份，赋形剂 80-160份，填充剂 80-120份，崩解剂30-50份，粘合剂30-60份，润滑剂5-15份。

2.根据权利要求1所述西罗莫司纳米片剂，其特征在于：所述西罗莫司纳米药物组合物中，西罗莫司纳米颗粒的平均粒径小于400nm，西罗莫司药物有效成分占15％-20％。

【产品名称】通用名称：甲状腺球蛋白检测试剂盒(电化学发光法)英文名称：Tg II【包装规格】100测试/盒【预期用途】主要用途用于体外定量测定人血清和血浆的甲状腺球蛋白含量。

Tg的测定能辅助监控甲状腺切除术后的情况。

Elecsys和cobas e 免疫分析仪的工作原理是电化学发光免疫分析“ECLIA”。

临床应用甲状腺球蛋白是一种分子量接近660 kDa的糖蛋白。

4Tg由甲状腺细胞大量合成并释放到甲状腺滤泡腔内。

TSH、甲状腺内碘缺乏、促甲状腺激素免疫球蛋白均可刺激Tg的生成。

Tg对外周甲状腺激素T3和T4的合成起决定作用。

它含有约130种酪氨酸残基，其中一部分在TPO(甲状腺过氧化物酶)和碘化物存在时被碘化为单碘-和二碘酪氨酸(MIT和DIT)。

3随后也是在Tg和TPO的作用下，MIT和DIT偶联结合成T3和T4。

5甲状腺细胞合成Tg以及将Tg转送到滤泡的过程中，少量蛋白可进入血流。

因此无甲状腺疾病的健康个体中也能检出低浓度Tg。

Tg浓度升高在不同的甲状腺疾病中均有报道，桥本氏病、葛瑞夫兹病等。

Tg还有助于鉴别亚急性甲状腺炎和人为甲状腺毒症。

对于先天性甲状腺功能减退症Tg的检测可用于鉴别先天性甲状腺缺失和甲状腺发育不全或其它病理情况。

5,6,7Tg检测主要用于甲状腺全切或次全切术后病人的随访。

由于甲状腺是Tg的唯一已知来源，在甲状腺全切或次全切伴随放射性碘成功消融残留甲状腺组织后，血清Tg浓度将降至极低，甚至检测不到。

对于部分甲状腺切除的病人，检测到的Tg水平取决于手术后残留的甲状腺组织的多少。

若甲状腺全切术后仍可检出Tg则提示DTC残留或复发。

因此Tg明显升高常提示该疾病复发。

8,9,10,11,12,13采用高灵敏度Tg检测后，“甲状腺球蛋白阳性”病人数量增加，即使这些病人并未表现出疾病症状。

13不能认为这些病人没有疾病，应根据当前指南进行监测随访。

已有不同临界值报道用以鉴别仅需监测的病人和那些需要接受进一步诊断和治疗的复发病人。

与4h 结果无显著性差异(P >0105),这可能是由于加封胶塞使试管内形成高压、高温所致。

日常工作采用高压水解或煮沸水解均可。

用碳酸盐缓冲液配制乙酰丙酮溶液可保证乙酰化时pH 的稳定。

据报道原法37°C 60m in 完成显色反应,但我们实验表明37°C 10m in 既可完成。

本文改进的方法使实验时间由原来的6～8h 缩短为1h ,更适于日常工作需要。

所测得的胃病患者胃黏膜组织中氨基己糖的含量与已往报道基本一致[1]。

改进后的方法仍未克服原法的部分缺点。

我们拟用胰蛋白酶水解,新的乙酰化试剂乙酰化,使反应能在中性条件下一步完成,目前正在进一步研究。

参考文献:[1]　徐肇敏,张志宏,陈载融,等1幽门螺杆菌破坏胃黏膜屏障的研究[J ]1中华内科杂志,1995;34(9):6031[2]　钱　伟,侯晓华,谢小平,等1消化性溃疡与黏膜中糖蛋白含量的关系[J ]1中华消化杂志,1993;13(4):2321[3]　刘海军,许国铭,李兆申1测定氨基己糖和磷脂含量对胃黏膜防御机制的评价[J ]1解放军医学杂志,1996;21(6):4501[4]　N euhaus OW ,L etzring M.D eterm inati on of hexo sam ines inconjuncti on w ith electropho resis on starch [J ].A nal Chem ,1957;29(8):12301[5]　张世尼,吴孟超1多种中性糖.氨基己糖及乙酰氨基己糖的微量测定[J ]1第二军医大学报,1990;11(5):4501[6]　王　军,李云华1用组合导数分光光度法同时测定氨基葡萄糖和氨基半乳糖[J ]1临床检验杂志,1991;9(2):771收稿日期:2000210219罗氏2010型电化学发光试剂残量的使用方法介绍Ξ陈光辉,程廷声,刘洁鸣　(广东省中山市小榄人民医院检验科,广东中山　528415)中图分类号:R 446　文献标识码:B 文章编号:100521716(2001)042024201 罗氏2010型电化学发光全自动免疫分析仪,采用先进的电化学发光技术,具有灵敏度高、线性范围宽、检测速度快,甚至可以检测核酸物质,测定试剂为专用试剂,2～5°C 可以稳定长达2年等优点,深受用户的欢迎。

罗氏电化学发光化学发光检测结果1.罗氏电化学发光是一种常用的实验技术。

Roche electrochemical luminescence is a commonly used experimental technique.2.这种化学发光技术可以用于快速检测血液中的生物分子。

This chemical luminescence technique can be used for rapid detection of biomolecules in blood.3.电化学发光方法可以提供高灵敏度和高特异性的检测。

Electrochemical luminescence methods can provide high sensitivity and high specificity detection.4.利用化学发光技术，可以进行多种生物标志物的定量分析。

Quantitative analysis of various biomarkers can be performed using chemical luminescence technology.5.这种检测方法不需要昂贵的设备，适用于基层医疗单位。

This detection method does not require expensive equipment and is suitable for primary medical units.6.电化学发光检测是一种快速、准确的方法。

Electrochemical luminescence detection is a fast and accurate method.7.高灵敏度的化学发光技术使得微量样品也能被可靠检测。

The high sensitivity of chemical luminescence technology allows reliable detection of trace samples.8.罗氏电化学发光系统结合了现代生物技术和光学原理。

罗氏电化学发光免疫分析仪测试项目定标物保存方法无信息定标物和质控物是测试包装内的一部分定标必须被进行：每盒新试剂；每天 (当仪器上使用同一盒试剂)；需要时，如质控不在范围内心肌标志物性激素项目肿瘤标志物贫血诊断指标、骨标志物传染病项目其它唐氏筛查先兆子痫罗氏电化学发光免疫分析仪定标物保存方法（简版）定标物不需要分装冰冻保存的项目：使用时从2-8︒C取出，摇均后吸200μl（此数据为国内实验室试验所得，不代表罗氏官方）到日立杯，马上将剩余定标品放回2-8︒CFT3、FT4、T3、T4、TSH、T-uptake、Anti-TG、HCG+β、HCG STAT、ProgesteroneCEA、CA 125、CA 153、CA 724、Cyfra21-1、NSE、Free PSA、HE4Myoglobin、Myoglobin - STAT 、Digoxin、Digitoxin、Ferritin、IgE、β-CrossLapsAnti-HAV、Anti-HAV IgM、HBsAg、HBsAg II quant、 Anti-HBs、HBeAg、Anti-HBe、Anti-HBc、 Anti-HBc IgM、 HIV combi、HIV Ag 、HIV combi PTToxo IgG、 Toxo IgM、Rubella IgG、Rubella IgM、CMV IgG、CMV IgM定标物需要分装冰冻保存的项目：定标物加水复溶，用子弹头进行分装（分装量为200μl（此数据为国内实验室试验所得，不代表罗氏官方）），然后放在-20︒C保存，使用时取出并平衡到室温TG、Anti-TPO、 Anti-TSHR、CK-MB、CK-MB STAT、ProBNP、Troponin T HS、Troponin T HS STAT free βHCG、PAPP-A、 Insulin、C-peptide、IL-6 、PCT、Anti-CCPCortisol、ACTH、Estradiol II、FSH、LH、Prolactin、Testosterone、DHEA-S、SHBG、PIGF、 sFlt-1、hGHAFP、CA 199、TPSA、S100Vitamin B12、Folate III 、N-MID、Total P1NP、PTH、PTH STAT、PTH(1-84)Vitamin D3、Vitamin D totalHSV-1 IgG、 HSV-2 IgG11 / 11。

【产品名称】通用名称：他克莫司检测试剂盒（电化学发光法）英文名称：Tacrolimus【包装规格】100测试/盒【预期用途】主要用途主要用于定量测定人全血中他克莫司的含量。

本检测有助于接受他克莫司治疗的心、肝、肾移植病人的治疗。

电化学发光免疫测定试剂“ECLIA”，适用于罗氏Elecsys 和cobas e免疫测定分析仪。

临床应用他克莫司（又称KF506）是一种大环内酯类抗生素，1984年在日本被发现，是链霉菌属的代谢产物1.2,3。

研究表明他克莫司的免疫抑制活性是环孢霉素的10-100倍。

4他克莫司产生免疫抑制效应的主要原理在于它可以抑制T细胞的激活和增殖。

细胞内的他克莫司可结合一种抑免蛋白FK506-结合蛋白（FKBP-12），这种免疫复合物可抑制磷酸酶的活性。

5抑制磷酸酶可限制激活的T细胞（NFAT）的核因子的去磷酸化和核转位，由此调节包括IL-2、IL-4、TNF-α和γ-干扰素在内的多种细胞因子的转录，进一步限制淋巴细胞的激活和增殖。

6,7,8,9,10他克莫司具有很高的亲脂性，吸收不完全并且不稳定。

吸收后，他克莫司高效结合蛋白和红细胞，血浆中99%的药物都与白蛋白或α-1-糖蛋白结合。

11他克莫司的生物利用度和代谢主要受细胞色素P450药物代谢同工酶CYP3A4和CYP3A5以及流出泵P-糖蛋白的影响，在表达和功能方面在同一个体以及不同个体之间都变现出明显的差异。

12,13,14他克莫司在同一病人和不同病人间都表现出很大的变异性，无论剂量高低都可能出现严重不良反应。

他克莫司浓度不足可能导致对抑制器官的排异反应。

浓度过高又可导致严重不良反应。

他克莫司的主要不良反应包括肾毒性、神经毒性、胃肠道损害、糖尿病、高血压病和恶性肿瘤。

15,16为了让每个病人的体内药物浓度都维持在狭窄的治疗窗范围内，治疗药物监测的应用和浓度控制下给药作为标准临床实践的一部分已在临床上应用多年，它也是病人治疗的主要手段。

××××第页共页1. 目的规范罗氏2010全自动电化学发光检测仪的操作程序,正确使用该仪器,保证检测工作的顺利进行及其结果的准确性和可靠性。

2. 适用范围此操作规程适用于罗氏2010全自动电化学发光检测仪的操作及维护3. 职责3.1 操作人员：必须按照本规程操作仪器进行相关项目的测定，并对仪器进行日常维护，作使用登记。

3.2 管理责任人：负责监督仪器的使用是否符合规程，设置仪器的参数，对仪器进行定期维护、保养。

3.3 科室负责人：负责该仪器的综合管理及解释。

4. 该SOP变动程序本标准操作程序的改动，可由任一使用本SOP的工作人员提出并报经下述人员批准签字：专业主管、科主任。

5. 操作程序5.1 检测前准备5.1.1 检查UPS电源、电脑主机与仪器的接口，检查打印纸是否足够。

5.1.2 检查系统试剂、系统水、tip头、cup杯等试剂、耗材是否充足，液体及固体废弃物是否清空，样品针和试剂针是否清洁无弯曲。

5.2 标本检测程序5.2.1开机：打开中文报告电脑开关及打印机开关，打开中文软件及接收程序，打开UPS电源及仪器保护罩，打开系统试剂盖（procell和cleancell）、仪器显示器电源及主机电源，等待仪器启动及自检。

5.2.2 仪器启动后，显示屏上显示：“”，用手指轻触显示屏登陆操作界面。

5.2.3 打开试剂仓盖（抓住把手向左旋至OPEN标志，然后移开），放入要测试项目试剂至1-15号试剂位，通用稀释液放在16-18号位置，放好试剂后，盖上试剂仓盖，并旋至LOCK 位置。

5.2.4 扫描试剂：INVENTORY→Reagent Scan 查看试剂的位置、试剂量，试剂是否需要定标（提示RC标志）及消耗品使用情况。

××××第页共页5.2.5 当仪器扫描完成后，屏幕上方显示Stand-by时，在样品盘1-30号位置依次放入待测样本，在最后一个样本后面插上贴有中止码信息的试管。