雅培生化大型生化仪上岗培训.

1、6 Sigma在临床生化检测的应用:Architect C16000 Kasai C, et al. Evaluation of general chemistry assays on the Abbott Architect C16000 clinicalM«q) XUCW30QCC- RqgMOI 一<Lambert-Beer!s Law彳亍生溶液对光的吸收程厦与吸光 成正比。这种关系称为朗冶比尔定律，其数学表达式为OA = Kbc c=A *1/kb F=KbA称光的强和I分别为入射光和透射 .通过的液层厚 /. r歷的樽，K为比例常数。b的单位为cm,窘c的箪忌攻mol

2、/L曩不，贝诵£表不光物光学检测原理光路系统为直接光检测，后分光，采用凹面衍射光栅，有16个检测波长。光路系统主要组成有：卤鹄灯，隔热玻璃，凸镜，Shutter,狭缝，反光镜，光栅，线性硅二极管矩阵，前置放大板，数据处理板和CPU板等。光路组件将光聚焦后射入反应应时，光栅后 I ;矩阵根1双波长杯，宀一亠被分， 据不 测定每个读数点，一般分初项目都使用双 沁上込训|关于反应时间反应时间包括反应杯 转盘的转动，转动的 时间和转盘周围组件 的位置每次转动都发生在特 定的时间和到达特定 的位置反应转盘按逆时针方 向每4.5秒旋转近1/4 圈（41个反应杯的位 置）使反应杯到达特 定位置关于

3、反应的几个点1 起始位登，样品探针将样胡吸入反 应杯2. R1位置，试剂探针1将1试剂 （或样品稀释液）加入反应杯3. 无动作4混匀位置仁样品和1试剂（或样品 稀秤液被混匀45 反应杯经过光路系统被读取吸光 度值5.以上总共转了4个1/4圈共164个反应 杯位克，此位克为起始1号位趕的后 一个反应杯饷位克（总共有165个反 应杯）。加果棒品需稀释,在此位 置样品探针将稀释样品吸入放入1号 位登的反应杯。31 如此样品进行ICT检测，在此位置 被ICT系统的探针从反应杯吸入稀释 样品进行检测67. R2位肖鳥试剂探针2将2试剂加入反 应杯（此前时间为51分钟）68. 混匀位置2,样品/试剂1和试

4、剂2被 混匀、几匕牛知诒as? a时nz1 24Sample R1 (blank reading)67)I68IReaction (sample R1* R2)132136AU)reagent 1Ifl9“ I4.95-reagent 2IJ/ :EW |扁 n4.8*optic read®first二wash start 丄blank read optionrreact, readtimc / flex rate read optionJCuvette position:8 二 uod p 虽卜二 uod peeRulodREACTION TIME TABLETotal react

5、ion time = 9.9 min. (same for each assay)WI21BA9 VPPO<4读数点示忌图aoueqosquApprsdmalely d1 minutesApproximately 9.6 minutes1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 10 17 18 19 2021 22 2324 25 23 27 28 29 30 3132 33 Photometric fpointSample and first reagent dispenseSecond reagent dispenseFirst mixSecond mi

6、x雅培诊断仪器详述9分配组件加液量9光路9反应时间9常见报警enwevF-CHE iaiisiba9 VPPO<<分配组件加液量详述Sample Volume 样本体积：DS Vol = Diluted sample Volume:稀释样本Reagent 1 Volume R1 试剂量： Reagent 2 Volume R2 试剂量: Smartwash Volume智能冲洗洗液量 Water/Diluent Vol for Cone. Reage nt: 總祐乱侧辰的鑼液的量 反应杯最小量及最大量Volume Toleran8：Volume range MirVMax Rang

7、e Precision20 to 50 pL nominal +/ 5%</- 2 %51 to 345 pL nominal +/ 2.5%</» 0.5%范围步进2 to 35 pL0.1 pL steps2 to 15 pL0.1 pL steps20 to 345 pL1.0 pL steps20 to 345 pl1 pL steps10 to 345 pL1.0 steps*0 to 345 pL1.0 pL steps*Min. 160 pl Max. 360 plNot including overaspiration volumeWater- and r

8、eage nt volume en tered determine the dilution ratiowizibaS VPPO<<光路详述Specificati onsOptions波长340 一 380 一 404 一 412 -444-476 -16500 一 524 一 548 一 572 一 604 -628光径660 - 700 - 748 804测量体积：5 mmMono-/Bichromatic(单或双)读数点：Min. 160 pLMono Reag.: 1-33 pointsMax. 360 pLBi Reag.:1-16 (Blank)吸光度范围17-33(R

9、eaction)吸光度线性1-33(每 18 秒) 0.1 to 3.0 AbsCFlUiCVf ettEWiaiK人S VPPOfi项目反应类型>终点法：End 一 upEnd 一 down> 速率法(Kinetik):Rate 一 upRate downeHMteW-eHE WISIBAS VPPOI4友应类型-终点法7终点法：»反应达到平衡»单双试剂都可以使用双试剂推荐使用Self blank反应类型-锲巨注2a>0UBq0sqvTotal Abs -Self blank Abs = A Abs未知样本浓度= Abs x cal FactorA Ab

10、sToia-Absself blank可读点范I耳Self blank AbsT 2 3 4 5 6 7 6 9 10 11 12 13 14 15 16J7 16 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 Sample DispenseITime R1R2 addition二Reaction StartCIMICVCHEIAII21KA9 VPPOff友应类致终点法2aoueqjosqva待测物浓度在定标的基础上进行计算e+evF-eHE wiziba9 VPPO<<e+evF-eHE wiziba9 VPPO<<反应类

11、型-速率法2e+evF-eHE wiziba9 VPPO<<e+evF-eHE wiziba9 VPPO<<aode-elosqvFor Enzymes 一因子用于计算未知待测物浓度=A Abs变化/min x Enzyme Factor or A Abs 变化/ sec x Factor16 17 R2 addition =Reaction StartSample DispenseTimeAbs reading pointse+evF-eHE wiziba9 VPPO<<雅培诊断enWCVF-eHE WI21BAS VPPOflenWCVF-eHE WI2

12、1BAS VPPOflFlex Read TimeMain Read Time正常样本Abs Range() o o o () () o41 Substrate Depletion髙浓度或高活性样本Flex Time Reading酣的线性扩展9样本稀释eFHttCVF-e w 121KA9 VPPOK速率法 FLEX TIME READING弹性速AbsorbaC"enWCVF-eHE WI21BAS VPPOflenWCVF-eHE WI21BAS VPPOflPhotometric PointsenWCVF-eHE WI21BAS VPPOfl样本稀释加样本(2.0 to 35

13、.0 pL)用来稀释的 反应杯用量反应的 反应杯加入秤释液 (10to 345 pL4.5 s x 1 cycle4.5s x 1 cycle加入稀算后的样本(2.0 to 15.0 pL)eFW+CVF-eHt WI21KAa vppoif样本稀释ExampleReagent volume:Water volume:R128252R228Dilution nameSampleDiluted sampleDiluentWaterDilution factorUndiluted4.01:1.001:22.01:1:991:510.08.0901:5:06Highest calibrator4.0

14、e+C¥F-eH£ WI21KAS VPPOI4Dilutio n 1 - Un dilutedOD =1riingS = (4.0 + 28 + 252 + 28 + 0) / 4.0 = 78SMax = (4.0 + 28 + 252 + 28 + 0) / 4.0 = 78Sample dilution factor = 1 * 78 / 78 = 1.00Dilution 2 1:2:OD = 1S = (2.0 + 28 + 252 + 28 + 0) / 2.0 = 155SMax = (4.0 + 28 + 252 + 28 + 0) / 4.0 = 78S

15、ample dilution factor = 1 * 155 / 78 = 1.99Dilution 3-1:5:OD = (10.0 + 90 + 0)/10.0= 10S = (8.0 + 28 + 252 + 28 + 0) / 8.0 = 39.5SMax = (4.0 + 28 + 252 + 28 + 0) / 4.0 = 78Sample dilution factor = 10 * 39.5/ 78 = 5.06/ < W121BA9 VPPOff空白选择Reagent Blank 试剂空白(R1 + R2 + Water as Sample)9 Self - Blan

16、k(R 1 and Sample)enwevF-ewE iaiisika9 VPPOffenwevF-ewE iaiisika9 VPPOffenwievfpHE wisibaS VPPOfienwevF-ewE iaiisika9 VPPOffenwevF-ewE iaiisika9 VPPOff终点法/速率法-试剂空白9定标过程使用今R1 + R2 + (Sample)=水，生理盐水或定标液AenwevF-ewE iaiisika9 VPPOffenwevF-ewE iaiisika9 VPPOffoueqosqu错误代码报警解释Blank Absorbance RangeBLK一个或多个

17、空白定标的吸光度超出BLK 限定的范围9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 2S 26 27 26 29 3C 31 32 33PhotometricPointsSecond Reagent Dispense Second MixenwevF-ewE iaiisika9 VPPOffwod.aZQUJOloud ec ocSCMX一sPUOO0S9suod.2G wo6BeCEpuooes 03 卜 l|9LCHx-z-2 $uod<22uo6«3<J 6±J 0sua)d.2G0Q.ESMuem±

18、;:assq<vU.2QOO)E 匚 pemH.EesaoueqjosqvModel A 皿YHT2 一 Mlijhoaos匚o6urEr(5匚 uo 匸 eo二 daQ oo定标的报警设置7个检测功能：今试剂空白吸光度检测9定标重复次数间的离散度检测9斜率检测9 Factor Comparison Check *9 SD检测Monotonic Check *Approximation Convergence ErroreHWVF-eHE IAII21BA9 VPPOff终点法/速率法-试剂空白9定标过程使用> R1 + R2 + (Sample) = Water, Saline

19、or Calibrator A错误代码报警解释Blank Absorbs nee RangeBLK一个或多个空白定标的吸光度超出BLK 限定的范围a ouraqosqvBLK Abs Range1 2 3 4 567 6 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 X 31 32 33enwevF-ewE iaiisika9 VPPOifSecond Reagent Dispense Second MixPhotometricPointsenwevF-ewE iaiisika9 VPPOifAbsorbance定徐

20、一离散度检测代码解释Calibrator absorbance rangeDEV定标几次重复超出限定ConcentrationenwtcvFeRE w 12丄 b人9 VPPOff宗标一斜率检测Absorbance错误代码e : opec s c kSPN 一空浓围一min. & max. allowed SpanConce ntration牛ri.e. between B ankand Ca 1BlankCFrtWeVFG+E WI21BAa vppoti7COMGbofut u«M|AcoucW bo".coucCIH1CWCHEWI21BAS VPPOff定标

21、设置一体积2Outline I BaseAssay Configuration - CocainCALIBRATION I QCI Smart WashRerun rulesCalib. Mod字Urwar | |Blank / Calib Replication Extrapolation%Interval (H)BLK:C1C2C3C4C5C6C7C8SampleIJPLLLLLLL趾LLLLLL匚L一氐 LLLLLLLLLBlank abs Range I 0.0 fo.oCal DeviationI 0.0OKDeleteLot Change ICtg. ChangeIIVOver I

22、nterval2-PointBLK-厂1-Polnt定标方法定标推荐：9在手册上没有官方的明确限定9到定标周期或换批号时，建议全线定标今换盒进行空白定标选项：Blank1- Point2- Point9 FullC"lCWCHEWI21BA3 vppoix定标设置自动稀释AssayConfiguratiorvOROSOOullww I Base | CALIBRATIONQCSmart WashRerun rulesOKInterval (H)Extrapolation%6 VolBLK：C1C2C5C6C7Lot ChangeCalib. ModeSpline | |Ctg. Ch

23、angeI llOver IntervalI IPI0.0Blank obo Range I 0.0| 0.0BLK.DG.Vol2-PointBLKBlank/Calib. ReplicationI 3/GomplcCal DeviationI 0.0HO I 20,0I 35.0I 35.0ll0.0CancepinSpan abs Range I 0.0W.Vol1-PointPrinttImportExportUpdateDelete定标设置自动稀释CIHlCWCHEWI21BAa vppoffCC Application TrainingLevelNameSample volumeD

24、iluted sample volumeDiluent volumeWater volumeDilution factorEn tered cone.*BlankWater5.01:10Cal 1Chem52.05.0981:50500Cal 2Chem510.05.0901:10500Cal 3Chem520.05.0B01:5500Cal 4Chem52.51:2500Cal 5Chem55.01:1500etCVf=eHE WI21BAS VPPOI4ASSAY RESULT FLAGGING分析结果确认CHWeVF-CHE IAII21BAS VPPOI4REACTION VALIDI

25、TYCHECKS9终点法9速率法9非线性&终点法-AbsMaxVar 检测最终吸光度的变异性-Absorbance Limit检汎在读数窗内吸光度变化 的线性-Colour Correctio n-颜色矫正-Reaction Check检测免疫反应的前代效应 （抗原过剩）CIMlCbCHEIAII21EAS VPPOff终点法稳定性的检测S R1 Blank Read option pointl up to 17R2Main Read TimeEnd Stability = AbsMxVar代码解释Main Read TimeABSAbs超出(一01 -3.0)AbsMaxVarRF读

26、数波动Main Read Timo option pointPhotometric18 upt to 33PointseHMtCVF-eH£ WI21BAa vppo<<速率/终点一吸光度限定ErrorInterpretationAbs Limi tA#0Main/Flex Read time-none of the read points within defined abs limitsA#1Only 1 point within defined abs limitsA#2Only two points within defined abs limitsABSAbs

27、OOR (0.1 -3.0)CHMtCVF-ettE WI21BA9 VPPOff速率法粒测在读数窗内吸光度变化的线性AbsorbancePhotometric PointsReading TimeCalculation:灼严 X100%= rate of linearity %代码解释Linearit yRL%Main/Flex Read 读数 窗口内的点超出了以 下限定：前三点与后 三点的速率之比的范 围a vppo<<终点/速率颜色矫正3«nipleSevufidCalculation: (Abs Reagent 1 & Sample Blank) (Abs

28、 Reagent Blank)CFtWeW-eHE W121KA a vppofi终点/速率颜色矫正AborbafKeColor correction read timeMainread timeAsortance rangupper limitT J1 AClusted Absorbancerange upper limitReagent BlankNormal samptePhotometric Rzd Points颜色矫正-例子etC¥F-eHE WI21BA9 VPPO<<Interpretation超出前带反应限定etC¥F-eHE WI21BA9 VPPO<<电解质系统的检测原理电解质检测是测定样品中的电位。使用集成晶 片技术（ICT）来检测钠，钾和氯。ICT技术是 将固项的离子选择电极组合在一个单一的晶片上，从而减少了在进行电极检测中所需要的保 养程序。ICT系统包括一个ICT的吸样针和一个ICT的模块。 品并送入IG 热的参比液 果的计算）1厂丁口&q