體外診斷試劑盒線性范圍研究線性區(qū)間評價(jià)資料及可報(bào)告區(qū)間建立

體外診斷試劑盒線性范圍研究/線性區(qū)間評價(jià)資料及可報(bào)告區(qū)間建立,,,,,,,,,

實(shí)驗(yàn)名稱,試劑盒線性范圍研究/線性區(qū)間評價(jià)及可報(bào)告區(qū)間建立,,,,,,,,

實(shí)驗(yàn)?zāi)康?確定試劑盒的線性范圍/線性區(qū)間可報(bào)告區(qū)間上限,,,,,,,,

實(shí)驗(yàn)方法,"依據(jù)體外診斷試劑分析性能評估系列指導(dǎo)原則(征求意見稿)以及體外診斷檢測系統(tǒng)性能評價(jià)方法第4部分：線性區(qū)間與可報(bào)告區(qū)間

基本要求

（1）樣本基質(zhì)應(yīng)與臨床實(shí)驗(yàn)樣本相似，但不可采用含有對測定方法具有明確干擾作用物質(zhì)的樣本，如溶血、脂血、黃疸或含有某些特定藥物的樣本。進(jìn)行血清學(xué)標(biāo)志物檢測時(shí)，理想的樣本為分析物濃度接近預(yù)期測定上限的混合人血清。

（2）建立一種定量測定方法的線性范圍時(shí)，需在預(yù)期測定范圍內(nèi)選擇7-11個(gè)濃度水平。如將預(yù)期測定范圍加寬至130%，在此范圍內(nèi)選擇更多的濃度水平，然后依據(jù)實(shí)驗(yàn)結(jié)果逐漸減少數(shù)據(jù)點(diǎn)直至表現(xiàn)出線性關(guān)系，可發(fā)現(xiàn)最寬的線性范圍。

（3）當(dāng)對標(biāo)稱線性參數(shù)進(jìn)行驗(yàn)證時(shí)，需在已知線性范圍內(nèi)選擇5-7個(gè)濃度水平。

（4）無論是建立或驗(yàn)證線性范圍，所選用的濃度水平應(yīng)可覆蓋整個(gè)預(yù)期測定范圍并包括與臨床有關(guān)的重要評價(jià)濃度，如最小測定濃度或線性范圍的最低限、不同的醫(yī)學(xué)決定水平、最大測定濃度或線性范圍的高限等。",,,,,,,,

說明,公式已在excel中設(shè)定好，在紅色區(qū)域錄入數(shù)據(jù)即可自動計(jì)算出結(jié)果，最終結(jié)論的得出需三批數(shù)據(jù)。,,,,,,,,

標(biāo)本配制：,,,,,,,,,

11個(gè)濃度水平的樣本制備,,,,,,,"建立線性范圍：需測定9-11個(gè)濃度水平，每個(gè)濃度水平重復(fù)測定3-4次。

驗(yàn)證標(biāo)稱線性參數(shù)：需測定4-6個(gè)濃度水平，每個(gè)濃度水平重復(fù)測定3-4次。

所有樣本應(yīng)在一次運(yùn)行中或幾次間隔很短的運(yùn)行中隨機(jī)測定，最好在一天之內(nèi)完成。",,

樣本號,1,2,3,4,5,6,,,

低濃度血清(ml),1.00,0.90,0.80,0.70,0.60,0.50,,,

高濃度血清(ml),0.00,0.10,0.20,0.30,0.40,0.50,,,

樣本號,7,8,9,10,11,,,,

低濃度血清(ml),0.40,0.30,0.20,0.10,0.00,,,,

高濃度血清(ml),0.60,0.70,0.80,0.90,1.00,,,,

5個(gè)濃度水平的樣本制備,,,,,,,,,

樣本號,1,2,3,4,5,,,,

低濃度血清(ml),1.00,0.75,0.50,0.25,0.00,,,,

高濃度血清(ml),0.00,0.25,0.50,0.75,1.00,,,,

檢測結(jié)果：,,,,,,,,,

樣本號,理論濃度,測定Y1,測定Y2,測定Y3,測定Y4,Y均值,Y標(biāo)準(zhǔn)差s,t1,t2

1,25.00,25.08,24.90,25.01,25.00,25.00,0.07,1.10,-1.33

2,42.50,42.48,42.49,42.60,42.59,42.54,0.06,0.90,-0.91

3,60.00,60.00,60.03,59.96,60.07,60.02,0.05,1.19,-1.15

4,77.50,77.48,77.59,77.44,77.56,77.52,0.07,1.02,-1.09

5,95.00,94.98,94.92,94.97,95.03,94.98,0.04,1.24,-1.19

6,112.50,112.47,112.42,112.59,112.45,112.48,0.07,1.45,-0.83

7,130.00,130.06,129.96,129.98,130.02,130.01,0.05,1.23,-1.08

8,147.50,147.44,147.52,147.58,147.49,147.51,0.06,1.24,-1.13

9,165.00,165.02,165.06,164.97,165.04,165.02,0.04,0.87,-1.40

10,182.50,182.48,182.48,182.54,182.59,182.52,0.05,1.30,-0.82

11,200.00,200.05,199.98,199.99,199.95,199.99,0.04,1.37,-1.02

t值臨界值,,,,,,,,,

樣本數(shù)(測定次數(shù)),顯著性水平,,,,樣本數(shù)(測定次數(shù)),顯著性水平,,,

,0.05,0.025,0.01,0.005,,0.05,0.025,0.01,0.005

3,1.153,1.155,1.155,1.155,4,1.463,1.481,1.492,1.496

如t1、t2值大于臨界值，則相應(yīng)的可疑值為離群值。,,,,,,,,,

"進(jìn)行多項(xiàng)回歸分析

對數(shù)據(jù)組進(jìn)行多項(xiàng)回歸分析，得到一級、二級與三級多項(xiàng)式。一級多項(xiàng)式為直線，二級多項(xiàng)式表示上升曲線或下降曲線，三級多項(xiàng)式表示S形曲線（在測量范圍兩端具有明顯的非線性）。

多項(xiàng)式方程如下：

級數(shù)多項(xiàng)式回歸自由度（Rdf）

一級Y=b0+b1X2

二級Y=b0+b1X+b2X23

三級Y=b0+b1X+b2X2+b3X34

3.5對回歸方程進(jìn)行線性檢驗(yàn)

多元回歸方程中以bi表示的系數(shù)為回歸系數(shù)。在二級與三級方程中，b2與b3為非線性系數(shù)。對回歸方程進(jìn)行線性檢驗(yàn)就是對每個(gè)非線性系數(shù)作t檢驗(yàn)，判斷回歸系數(shù)與零是否有顯著性差異。b0與b1不反映非線性，故不需對其進(jìn)行檢驗(yàn)。對b2與b3的檢驗(yàn)方法如下：

計(jì)算統(tǒng)計(jì)量t，計(jì)算公式為：t=bi/SEi

其中，SEi為每個(gè)非線性系數(shù)的斜率標(biāo)準(zhǔn)誤，計(jì)算公式為：

其中，Y為回歸方程預(yù)測值，與為測定均值。",,,,,,,,,

,,,,,,,,,,

回歸分析結(jié)果,,,樣品數(shù)：,11,測定次數(shù),4,,,

回歸方程,方程因子,方程因子值,SEi,t-test(t=bi/Sei）,標(biāo)準(zhǔn)估計(jì)誤差Sy.x,自由度df=L?R-Rdf,最適方程,結(jié)果分析,

"一次方程

(y=b1X+b0)",b0,-0.01,0.01,-0.82,,,,,

,b1,1.00,0.00,9002.81,0.02,42,,,

二次方程(y=b2X2+b1X+b0),b0,-0.02,0.03,-0.91,,,,,

,b1,1.00,0.00,1838.56,,,,t0.05(ν)=,2.020

,b2,0.00,0.00,-0.57,0.02,41,否,小于，無統(tǒng)計(jì)學(xué)意義,

三次方程(y=b3X3+b2X2+b1X+b0),b0,-0.04,0.05,-0.80,,,,,

,b1,1.00,0.00,549.10,,,,,

,b2,0.00,0.00,-0.46,,,,t0.05(ν)=,2.021

,b3,0.00,0.00,0.39,0.02,40,否,小于，無統(tǒng)計(jì)學(xué)意義,

,,,,,,,,,,

"臨床標(biāo)準(zhǔn)的線性檢驗(yàn),如回歸分析滿足要求，則可省略",,,,,,,,,

如ADL＜5%，則擬合的多項(xiàng)式無臨床意義,,,,,,,,,

樣本號,測量均值,一次方程預(yù)期值,二次方程預(yù)期值,三次方程預(yù)期值,二次與一次絕對偏倚,二次與一次相對偏倚%,三次與一次絕對偏倚,三次與一次相對偏倚%,

1,25.00,24.99,24.98,24.98,-0.01,-0.025%,-0.01,-0.041%,

2,42.54,42.53,42.53,42.53,0.00,-0.006%,0.00,-0.004%,

3,60.02,60.01,60.01,60.01,0.00,0.001%,0.00,0.006%,

4,77.52,77.51,77.51,77.51,0.00,0.003%,0.01,0.007%,

5,94.98,94.97,94.97,94.97,0.00,0.004%,0.01,0.006%,

6,112.48,112.47,112.48,112.48,0.00,0.004%,0.00,0.004%,

7,130.01,130.00,130.00,130.00,0.00,0.003%,0.00,0.001%,

8,147.51,147.50,147.50,147.50,0.00,0.002%,0.00,0.000%,

9,165.02,165.02,165.02,165.02,0.00,0.000%,0.00,-0.002%,

10,182.52,182.52,182.52,182.52,0.00,-0.001%,0.00,-0.002%,

11,199.99,199.99,199.98,199.98,-0.01,-0.003%,0.00,-0.001%,

從上表數(shù)據(jù)判定絕對偏倚和相對偏倚是否滿足要求（法定或自定義標(biāo)準(zhǔn)）。,,,,,,,,,

可報(bào)告區(qū)間建立（本標(biāo)準(zhǔn)僅針對上限，下限為定量限）,,,,,,,,,

高值樣本號,理論濃度,樣本稀釋比例,重復(fù)測量結(jié)果1,重復(fù)測量結(jié)果2,重復(fù)測量結(jié)果3,實(shí)測均值,還原濃度,相對偏倚,

樣本1,2000,5,387.6,387.6,416,397.0666667,1985.333333,-0.73%,

,,25,75.352,79.384,76.752,77.16266667,1929.066667,-3.55%,

,,50,37.552,43.348,38.308,39.736,1986.8,-0.66%,

,,100,22.002,18.856,21.342,20.73333333,2073.333333,3.67%,

,,200,8.739,8.818,8.689,8.748666667,1749.733333,-12.51%,

樣本2,1800,5,366.12,359.1,362.52,362.58,1812.9,0.72%,

,,25,72.1512,71.748,70.236,71.3784,1784.46,-0.86%,

,,50,35.676,36.486,36.4212,36.1944,1809.72,0.54%,

,,100,16.9506,19.8018,19.1484,18.6336,1863.36,3.52%,

,,200,8.0413,7.9766,7.7254,7.914433333,1582.886667,-12.06%,

樣本3,1600,5,309.28,311.68,324.32,315.0933333,1575.466667,-1.53%,

,,25,62.4768,61.5808,66.1504,63.40266667,1585.066667,-0.93%,

,,50,34.0448,33.7568,34.8224,34.208,1710.4,6.90%,

,,100,16.8624,14.7152,14.7328,15.4368,1543.68,-3.52%,

,,200,6.5512,7.0936,7.1144,6.919733333,1383.946667,-13.50%,

以相對偏倚不大于設(shè)定偏倚為判斷標(biāo)準(zhǔn)，選取三份樣本的相對偏倚均不大于設(shè)定偏倚的最大稀釋倍數(shù)為方法推薦的最大稀釋倍數(shù)（小于此倍數(shù)的三份樣本所有稀釋比例的相對偏倚均不大于設(shè)定偏倚），測量區(qū)間上限與最大稀釋倍數(shù)的乘積為該方法可報(bào)告區(qū)間的上限。,,,,,,,,,

假定10%為偏倚上限，則3份樣本在稀釋倍數(shù)100倍及以下時(shí)，實(shí)測均值與理論值的偏倚均小于±10%，而200倍時(shí)兩份樣本偏倚已超過允許偏倚，故本項(xiàng)目最大稀釋倍數(shù)為100。,,,,,,,,,

ADL計(jì)算：,,,,,,,,,

內(nèi)容,,,二次方程,三次方程,,,,,

指導(dǎo)原則方法ADL=,,,0.008%,0.006%,,,,,

行標(biāo)方法ADL中間計(jì)算值,,,0.0006,0.0009,,,,,

行標(biāo)方法ADL=,,,0.003%,0.004%,,,,,

非線性臨界值PctBnd=,,,5%,5%,,,,,

行標(biāo)公式,,,,指導(dǎo)原則公式,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,p(x):最優(yōu)擬合二階或三階方程的擬合值,,,,

,,,,,b0+b1x：擬合一階方程的擬合值,,,,

對數(shù)據(jù)進(jìn)行精密度檢驗(yàn)，線性評價(jià)前提,,,,,L：樣本數(shù),,,,

,,,,,ˉc=（y1+y2+y3+……+yn）/n,,,,

指導(dǎo)原則公式,,,行標(biāo)公式,,n：樣本數(shù)乘以重復(fù)次數(shù),,,,

,,,,,d:最優(yōu)擬合方程的階,,,,

,,,,,σ：最優(yōu)擬合發(fā)能給出的回歸標(biāo)準(zhǔn)誤,,,,

,,,,,Rdf:回歸自由度,,,,

,,,,,R：重復(fù)測量的次數(shù),,,,

,,,,,C：常數(shù),,,,

,,,,,,,,,,

樣本號,（yi-（Y一階回歸））^2,（yi-（Y二階回歸））^2,（yi-（Y三階回歸））^2,SDr計(jì)算中間值,CVr計(jì)算中間值,,,,

1,0.0000,0.0096,0.0061,0.0165,0.0000,,,,

2,0.0123,0.0021,0.0016,0.0119,0.0000,,,,

3,0.0076,0.0001,0.0005,0.0072,0.0000,,,,

4,0.0139,0.0011,0.0051,0.0136,0.0000,,,,

5,0.0057,0.0001,0.0024,0.0055,0.0000,,,,

6,0.0161,0.0001,0.0034,0.0159,0.0000,,