外文翻译-外文文献-英文文献-生化的分析者在将Olli-C---D使用在朝派酶学一年后的评价

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1、 Evaluation of the Olli C + D biochemical analyser after over a year of use in enzymologyG. Baraton, D. Grafmeyer, A. Capuano, L. Richard and J. Sofia.Laboratoire automatisde Biochimie Clinique, (Professeur J. Bertrand), HopitalEdouardHerriot, Place dArsonval, 69374 Lyon Cedex 2, FranceTo meet today

2、s clinical requirements, clinical biochemistry laboratories must be increasingly automated. The required criteria for an automatic analyser are reliability; the ability to change chemical reactions easily and quickly; rapid determinations and low analysis cost.It appears that the Olli C + D parallel

3、 analyser (Kone Oy instrument division Espoo, Finland) answers these criteria. A report on the Olli C system has already been published by Puuka and Pukka 1. This evaluation, using the C + Dsystem, deals with its intensive and daily use in enzymology over a period lasting more than one year.Descript

4、ionThe instrument is a parallel, multichannel, photometric analyser. The different phases of determination are performed in batches of 24 samples including blanks, standards and controls. The rate of analysis is 700 tests per hour with end point determinations of 360 kinetic measurements per hour re

5、action time is 21A minutes.The analyser comprises three independent units, the sample diluter/processor, photometer (each controlled by microprocessor) and incubator. The sample change is performed by hand using disposable cuvettes in a thermostatedblock.The Olli D Diluter measures 350 x 1100 x 740

6、mm and weighs 70 kg. It is made up of a dispensing tray with 24 cuvette blocks which fit in the Olli C photometer, eight reagent vessels, one block with 24 serum vials, an eight tip dispensing head and a digital display and keyboard. The diluter can be programmed by hand or by tape cassette for 30 a

7、nalyses; it is possible to add serum and up to eight reagents distributed in one or several runs. Each analysis programme follows the sequence:l. Attain reaction temperature (25, 30, 37C). 2. Measure reagent (5 to 1000/al by steps of 1/.tl) and position. 3. Measure and add sample. 4. Mix(5 to 180 se

8、conds).The photometer measures 330 x 520 x 740 mm and weighs 48 kg. It comprises a computer, a thermal pri_nter and a thermostated measuring chamber (25, 30, 37C). The light source is a zenon lamp. A quartz optic fibre system allows the simultaneous measurement of 24 cuvettes. Filters have a bandwid

9、th of 8 to 15 nm. The smallest readable volume is 500 /al for end point determinations, and 300/1 for kinetic determinations.The photometer can be programmed by hand or by tape cassette for 15 analyses which may be made in end point mode, with or without blank measurement, or in kinetic mode, with o

10、r without reagent or serum blanks. Kinetics determinations are made using linear regression of 12 to 24 measurements in to 10 minutes. The results are printed with the name and units of the programmed analysis. Error messages are printed; eg, if the chosen initial absorbance limits have been exceede

11、d or if there is non-linearity of reaction. In addition, it is possible to have printed thetwelve or 24 absorbance measurements used for the calculation of the activity of each determination. The Olli incubator can handle four thermostated blocks. Each block contains one heating element and temperat

12、ure regulation is maintained via a water circulating system. The incubation period is determined by a timer and each block incubation time is terminated with an audible alarm.Material and metlodsThe evaluation was carried out according to the rules suggestedby the French Society for Clinical Biology

13、 (SFBC) 2, 3.Dilution and measurementAccuracy, within-block and day-to-day, was tested with a solution of drawing ink (Mars 745 from Staedler Germany) prepared as follows: 1.7 ml of ink as supplied was dissolved by gentle stirring in 1000 ml of distilled water and filtered in a buchner funnel throug

14、h silicone paper. The solution was preserved with Merseptyl and stored at +4C. Dilutions were made by the Olli D processor into Olli arcylic cuvettes and absorbances were read on the Olli C photometer at 405 nm. Measurements were verified by manual dilution followed by a reading using a Miniken spec

15、trophotometer (Coultronic).Control of temperatureThe control of the temperature variations between 25 and 37C was tested by measuring the absorbance of a paranitrophenol solution, (15 mg in 1000 ml of Tris HCL 0.2 M, pH 6.8) at 405nm, Naudin et al 4. The absorbances obtained at the chosen temperatur

16、e were compared with those obtained using four thermostated analysers; Miniken (Coultronic),Cobas BIO (Roche,) PA 800 (Vitratron), ACP 5040 (Eppendorf) On the Olli C, the time needed to reach the set temperature of 30C or 37C with an analysis volume of 550 ul(contained in the acrylic cuvette in the

17、thermostated block)was followed by noting the decrease in the absorbance of theparanitrophenol solution. The temperatures of cuvette contents were also monitored with a Metrix temperature probe (HA 1159).Optical performanceThis was tested according to the procedure of Rand 5.Linearity and precision

18、were tested using the following kits:UV-Trol (Biomerieux France) and Holnicob (Biotrol France).MethodAll analyses were performed at either 30C or 37C. The instrument settings were those recommended by the manufacturer.Glutamic oxaloacetic transaminase (SGOT) and pyruvic transaminase (SGPT) were meas

19、ured by the Wroblewski method at 37C or by the recommended method of the SFBC at 30C with the Biomerieux kits. Alkaline phosphatase (ALP) was measured following the recommended procedure of the German Society of Clinical Biochemstry (DGKC) at 37C. Accuracy, within day and day to day, and carryover w

20、ere tested using low, medium and high concentration control sera spiked where necessary with animal enzymes. These control sera were furnished by a local association operating in biological measurement quality control. (Pro Bio. Qual, Lyon, France).Results and discussionAccuracy and precision of the

21、 Olli D sample processor The results are presented in Table 1. Precision expressed as the coefficient of variation for 24 determinations was never greater than 0.925% for the most important dilution (10 of drawing ink in 510 1 of water).The measured absorbances varied directly with the amount of dra

22、wing ink added and were indistinguishable from those obtained by manual dilution. Day-to-day precision was 0.609% for the 1/12 dilution used routinely for SGOT and SGPT and 0.925% for the 1/30 dilution used routinely for ALP.The absorbance of drawing ink is temperature independant.The maximum variat

23、ion for a temperature range of 20-40C and an absorbance of 1.690 was + 0.002.Temperature controlWith the different automatic analysers the solution of paranitrophenol gave an average inverse ratio of 0.029 absorbance units for an increase of 1-C. With the Olli C, the absorbancesfound were 1.948 + 0.

24、002 at 25C and 1.595 + 0.004 at 37C, or 0.029 absorbance units/degree.The desired termperature was reached rather slowly. If the blocks and cuvettes were preheated to the designated temperature of 37 C, the usual volume of para-nItrophenol O (at the lmtlal temperature of + 4 C) reached 37 C in 8 min

25、utes in the Olli C and 7 minutes in the incubator (Figure 1).Similar results (not shown) were found for a set temperature of 30C, the steady state was reached in 6 minutes for the Olli C, and 5 minutes for the incubator.The block has to be manipulated outside the apparatus,and it is therefore impor_

26、tant to know the rate of cooling. At a set temperature of 37C, the block outside the apparatus cooled by 0.5C during the first minute and then C/minute during the following 5 minutes. Measurements were made with a temperature probe at ambient temperature.To avoid this cooling when the blocks were ma

27、nipulated outside the incubator for either the Olli C or the Olli D, the set temperature in the incubator was set slightly higher, at30.2 for 30C and 37.4 for 37C.Accuracy and precision of the photometer Linear absorption was maintained up to 2.2 units of absorbance at 340 nm and up to 2.8 at 405 nm

28、. Accuracy and precision, tested at 340 nm with the UV Trol kit (stabilized solution of NADH) were both good. Similar results at 660 nm were found with the Holnicob-kit (cobalt nitrate).Analytical results precisionWithin-day and day-to-day precision measurements, precision for SGOT measurements at 3

29、7C are presented in Tables 2 and 3. The CV did not exceed 2.1% at any of the levels of serum activity studied. The SGPT and ALP values,measured at 37C, (valued not shown) were similar. SFBC recommendations specify within-day precision for SGOT to be within 2.0% at the low level, 0.9% at the medium l

30、evel and 1.9% at the high level of activity.No carry over was detected when one row of eight high level control sera was followed by two rows of eight low level control sera.Table 2. Within day precision for SGOT measurements Results obtained using protocol A established by the SFBC.Day-to-day preci

31、sion and accuracySera with three levels of SGPT activity were assayed each month for one year. The results are shown in Figure 2 where each point represents two to five daily determinations.Similar results were found (not shown) for SGOT and ALP where the CV was never more than 7% at any considered

32、serum level.ReliabilityNo injury to operators was caused or seemed likely during the twelve month period of the test.Figure 2. Each point represents the average of two to five daily control determinations on sera with three levels of activity during a month. (*- low level;-medium level; high level).

33、Table 3. Between days precision for SGOT measurements Results obtained using protocol A established by the SFBC 2 3. The electronics performed well but were very sensitive to even a short power interruption, so the analyser should be electrically stabilised to prevent the Olli D or Olli C of the com

34、puter memory being erased. Systematic cleaning was carded out between each analysis and no problems were experienced with theneedle obstruction.Manual use of the keyboard and programming of the microprocessorOlli D and C was easy.ConclusionThe Olli C + D analyser with its incubator proved to be reli

35、able and sufficiently precise. Its advantages are speed, end point or kinetics measurements for enzymology or immunoenzymology,and the absence of immobilisation of the Olli C or D during long incubation periods.Each analytical step is displayed and many malfunctions and errors are indicated. Reliabl

36、e results were obtained over a period of more than one year with 300-600 daily enzymatic determinations. A block for reactions volumes of 300ul.d is being developed by Kone and should lower reagent costs.ACKNOWLEDGEMENTSThe authors wish to thank Mrs Bonnet Paulien for reading the manuscript and Mr P

37、anigoni (Kone Oy Instrument) for his help during this work.REFERENCES1 Puuka, R. and Puuka, M. (1980),JAutomat Chem 2, 153-1582 Collombel, C. (1977),Ana Biol Clin, 35,167-1923 Grafmeyer,D. (1978) Etude des performances globales dapres le protocole A S.F.B.C. in Societe Francaise de Biologie Clinique

38、.Mesure des Activites enzymatiques en Biologie Clinique.Grafmeyer, G., Collombel, C., Dingeon, B., Fournier, M., Mathieu,N., Varennes, J.P., Eds Association Amicale des Etudiants en Pharmacie, Lyon, FRANCE, pp 129-1374 Naudin, C. and Bailly, M. (1978), Etude de la thermostatation des analyseurs denz

39、ymes in Societe Francaise de Biologie Clinique. Grafmeyer, D., Collombel, C., Dingeon, B., Fournier,M., Mathieu, M., Varennes, J.P., Eds Association Amicale des Etudiants en Pharmacie, Lyon, FRANCE, pp 145-1475 Rand, R.N. (1969), Clin Chem, 15, 839-863生化的分析者在将Olli C + D使用在朝派酶学一年后的评价G. Baraton, D. Gr

40、afmeyer, A. Capuano, L. Richard and J. SofiaLaboratoire automatisde Biochimie Clinique, (Professeur J. Bertrand),HopitalEdouardHerriotPlace dArsonval, 69374 Lyon Cedex 2, France为符合现代临床的要求,临床的生物化学实验室一定是越来越要求对自动分析者提供可靠性的标准;容易和迅速改变化学反应的性能;快速决定和降低分析估计成本.例如Olli C+D平行分析者(Kone Oy工具部门Espoo,芬兰)回答这些标准.一些关于Oll

41、i C系统的报告已经已经被被Puuka出版和分量足的1.超过一年这评价处理它的仔细和每天使用在朝派酶学剩余物一时期耐久使用C+Dsystem.绪论：工具是一个平行,多信道的,光度学的分析者.决定反应的不同阶段履行在朝派批包含空白,标准和控制手段24样品.21A分钟分析的速度是根据根据小时反应时间存在360运动的度量的小时有的端点决定700个试验.分析者包括三个独立单位,样品稀释者/处理器,(每个被微处理机)和孵化器控制光度计.样品改变被手履行使用在一thermostatedblock中可处理的透明小容器.Olli D稀释者尺寸为350 x1100 x740毫米和重量是70千克.它存在编造的a,

42、用24透明小容器起跑器,其安排Olli C光度计,八艘试剂船,有24浆液小瓶的一起跑器,一配发头儿和一根手指八小费的配发盘子作炫耀行为和键盘.稀释者能被手为编制程序或者在附近磁带盒为30细察;添加浆液是可能的和多达八试剂分配在朝派一或者几跑步.每一分析程序随着sequence:l到来.达到反作用温度(25,30,37C).2.测量试剂(5比1000/al 1/.tl)的在附近步和位置.3.测量和添加样品.4.(5到180次品)混合.光度计尺寸为330 x520 x740毫米和重量是48千克.它包括一台电脑,一热的pri_nter和一温度自动调节器测量房间(25,30,37C).亮来源是一盏ze

43、non灯.一石英眼的纤维系统允许24个透明小容器的同时度量.过滤器有一8到15 nm的带宽.最小有趣容量是500/al为端点决定和300/1为运动的决定光度计能被手为编制程序或者在附近磁带盒为15细察哪一个可以被在随着或者没有空白度量端点方式中或者在运动的方式,有的金色外面试剂或者浆液空白中作.动力学决定被作10分钟使用在向中12到24度量的线的退回.结果被随着为分析编制程序的名字和单位加印.错误信息被加印;例如,如果选择开头吸光率极限已经被超过,金色条件有非线性的反作用.此外,已经印是可能的十二或者24吸光率度量为活动的计算使用每一决定.Olli孵化器能处理四温度自动调节器起跑器.每一起跑器

44、含有一电热元件和温度规则被经由一水保持传播系统.孵化期被一个计时员决心和每起跑器孵卵时间被随着一个听得见的警报结束.材料和方法：评价被按照被法国社会显示为临床的生物学(SFBC)2,3规定实行.稀释和度量：准确性在-起跑器以内-和天天被用一如下准备吸引墨水(火星745从Staedler德意志)溶液测验:如同提供那样1.7 ml的墨水被和蔼在1000年搅拌蒸馏水的ml解散和渗入一buchner漏斗通过硅氧烷纸.解决方案被用Merseptyl保持和在+4C储藏.稀释被Olli D处理器使变成Olli arcylic透明小容器和吸光率在405 nm被在Olli C光度计上阅读.度量被手工稀释查证偏于

45、随着一阅读到来使用一小视野分光光度计(Coultronic)对温度的控制：对温度变化在中间25和37C的控制被测量一paranitrophenol解决方案的吸光率,15在Tris HCL0.2米的1000 ml中mg,在405nm,Naudin etal 4pH值6.8测验.选择温度是的吸光率得到阿特把温度自动调节器分析者和得到使用四那些比较;小视野Coultronic共同灵魂传记Roche PA800 VitratronACP 5040Eppendorf在Olli C上时间需要随着被一550的分析容量ul包含在温度自动调节器起跑器was跟随的丙烯酸的透明小容器在朝派的达到30C或者37C的装

46、置温度在吸光率的中指出减少 paranitrophenol解决方案.透明小容器目录的温度被用一Metrix温度探测器(HA 1159)也监视.光学表现：这个被根据Rand5.Linearity的过程测验和精确性被测验使用下列的kits:UV-Trol(Biomerieux法兰西)和Holnicob(Biotrol法兰西).方法：所有的一切细察履行阿特不是30C就是37C.工具设置是被manufacturer.Glutamic草酰乙酸的转氨酶(SGOT)推荐那些在30C用Biomerieux配套元件和pyruvic转氨酶(SGPT)被按照在37C Wroblewski方法或者按照SFBC的推荐方

47、法测量.碱性磷酸酶(高山)was测量在37C随着临床的Biochemstry(DGKC)的德国社会的推荐过程到来.在白天和白天向白天和留下的部分以内准确性被测验用动物酶使用必然的地方,以钉状物固定低,中等和高集中控制手段浆液.这些控制手段浆液被一种在生物的度量质量控制控制中运作本地联系陈设.赞成Bio.Qual,里昂,法兰西.结果和讨论：结果在1表中展示的Olli D样品处理器的准确性和精确性.当为24决定协同因素的变化从不是为吸引在510 1的水中墨水中最最重要稀释10超过0.925%时,精确性表示.仔细斟酌吸光率用吸引墨水的总量外加的直接改变和是和被手工稀释得到那些无法区分.为1/12稀释

48、日常的精确性was 0.609%被为例行公事地用于ALP的1/30稀释例行公事地用于SGOT和SGPT和0.925%的吸光率吸引墨水存在温度的20-40C和一吸光率的1.690的independant.The最大变化为一温度范围是+0.002.温度控制手段：和不同自动分析者paranitrophenol溶液为一1-C的增加给出一平均的相反的0.029个吸光率单位比率.用Olli C吸光率裁决是1.948在25C和1.595+0.004在37C或者0.029吸光率单位度方面是 +0.002期望温度是相当缓慢达到.条件起跑器和透明小容器被预热向标示温度的37 C通常会发生的事情容量的帕拉-nItr

49、ophenol O(阿特lmtlal温度的+4 C)达到37 C在朝派8分钟在朝派Olli C和7分钟在朝派孵化器(图1).Similarresults(不展示)被发现为a设定温度的30C稳恒态was达到在朝派6分钟为Olli C和5分钟为孵化器.起跑器必须被在装置之外操纵,和它因此是impor_tant知道冷下来的速度.在一37C的装置温度方面,在下列的5分钟在装置之外起跑器经过0.5C在第一分钟然后C/分钟冷下来.度量是制做有的一温度探测器是在周围的温度方面避开这个当起跑器是的时候,冷下来在外面操纵为或者或者Olli COlli D孵化器,孵化器was略微高设定的装置温度在朝派,为37C

50、at30.2为30C和37.4.光度计线的吸收was的准确性和精确性在340 nm和多达2.8在405 nm保持吸光率的多达2.2个单位.在340 nm测验有UV Trol配套元件(使还原型烟酰胺腺嘌呤二核苷酸)溶液稳定的准确性和精确性是两个好处.在660 nm相似结果被用Holnicob-配套元件(钴硝酸盐)找出.分析的结果精确性：在-白天和日常的精确性度量以内-,-为在37C公亩SGOT度量精确性赠送在朝派桌子2和3.CV没有在浆液活动的学习水平任何方面超过2.1%.在37C测量,(,珍视未展示)SGPT和ALP价值观是相似.SFBC推荐在-白天精确性以内-为在低水平方面2.0%,中等的第

51、在方面0.9%和在高水平的活动水平方面1.9%级以内SGOT向是明确地讲.禁止射程剩余物was察觉什么时候被随排八高水平的控制手段浆液而来的是两排八低差不多控制手段浆液的.表2.在为SGOT度量白天精确性以内结果得到公认使用被SFBC建立礼仪A日常的精确性和准确性：每月随着SGPT活动的三水平浆液被分析在一年中.结果被向展示每一在哪里点代表二比五每天determinations.Similar结果的在朝派图2被发现未为SGOT和ALP,那里CV在任何考虑浆液水平方面从不是超过7%的展示.可靠性：在试验的十二月时期禁止对操作员伤害被造成或者似乎是可能.图2.在一个月每一点随着活动的三水平代表在浆

52、液上二比五每天控制手段决定的平均.*-低level;-中等水平;高水平的.表3.在为SGOT度量几天精确性之间结果得到公认使用被SFBC23建立礼仪A.电子学好性能但是是非常敏感是甚至一简洁力量中断,所以分析者应该用电力是stabilised阻碍电脑记忆存在的Olli D或者Olli C擦掉系统打扫从里面在每一分析之间被在上附卡片和没有problems是有经验用是theneedle障碍键盘和微处理机Olli D和C was的编程的手工使用是容易结论：它的孵化器证明Olli C+D分析者用是可靠和足够准确的.在长孵化期时期它的优势为Olli C或者D的酶学金色immunoenzymology和没

53、有不动化是速度,端点或者动力学度量.每一分析的步被展示和很多功能障碍和错误被指示.在一超过一年的时期的时间中可靠结果300-600每天酶的决定地被得到.一起跑器为反作用大量的300ul.d存在存在发展在附近Kone和是应该降低试剂costs致谢：作者希望感谢夫人女帽Paulien和先生Panigoni在这工作期间修改手稿 (Kone Oy他的帮助的工具.)参考文献：1 Puuka, R. and Puuka, M. (1980),JAutomat Chem 2, 153-1582 Collombel, C. (1977),Ana Biol Clin, 35,167-1923 Grafmeyer

54、,D. (1978) Etude des performances globales dapres le protocole A S.F.B.C. in Societe Francaise de Biologie Clinique.Mesure des Activites enzymatiques en Biologie Clinique.Grafmeyer, G., Collombel, C., Dingeon, B., Fournier, M., Mathieu,N., Varennes, J.P., Eds Association Amicale des Etudiants en Pha

55、rmacie, Lyon, FRANCE, pp 129-1374 Naudin, C. and Bailly, M. (1978), Etude de la thermostatation des analyseurs denzymes in Societe Francaise de Biologie Clinique. Grafmeyer, D., Collombel, C., Dingeon, B., Fournier,M., Mathieu, M., Varennes, J.P., Eds Association Amicale des Etudiants en Pharmacie, Lyon, FRANCE, pp 145-1475 Rand, R.N. (1969), Clin Chem, 15, 839-86314