體外診斷試劑盒線性范圍研究/線性區間評價資料及可報告區間建立,,,,,,,,,

實驗名稱,試劑盒線性范圍研究/線性區間評價及可報告區間建立,,,,,,,,

實驗目的,確定試劑盒的線性范圍/線性區間可報告區間上限,,,,,,,,

實驗方法,"依據體外診斷試劑分析性能評估系列指導原則(征求意見稿)以及體外診斷檢測系統性能評價方法第4部分：線性區間與可報告區間

基本要求

（1）樣本基質應與臨床實驗樣本相似，但不可采用含有對測定方法具有明確干擾作用物質的樣本，如溶血、脂血、黃疸或含有某些特定藥物的樣本。進行血清學標志物檢測時，理想的樣本為分析物濃度接近預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，需在預期測定范圍內選擇7-11個濃度水平。如將預期測定范圍加寬至130%，在此范圍內選擇更多的濃度水平，然后依據實驗結果逐漸減少數據點直至表現出線性關系，可發現最寬的線性范圍。

（3）當對標稱線性參數進行驗證時，需在已知線性范圍內選擇5-7個濃度水平。

（4）無論是建立或驗證線性范圍，所選用的濃度水平應可覆蓋整個預期測定范圍并包括與臨床有關的重要評價濃度，如最小測定濃度或線性范圍的最低限、不同的醫學決定水平、最大測定濃度或線性范圍的高限等。",,,,,,,,

說明,公式已在excel中設定好，在紅色區域錄入數據即可自動計算出結果，最終結論的得出需三批數據。,,,,,,,,

標本配制：,,,,,,,,,

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

驗證標稱線性參數：需測定4-6個濃度水平，每個濃度水平重復測定3-4次。

所有樣本應在一次運行中或幾次間隔很短的運行中隨機測定，最好在一天之內完成。",,

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標準差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(測定次數),顯著性水平,,,,樣本數(測定次數),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，則相應的可疑值為離群值。,,,,,,,,,

"進行多項回歸分析

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

多項式方程如下：

級數多項式回歸自由度（Rdf）

一級Y=b0+b1X2

二級Y=b0+b1X+b2X23

三級Y=b0+b1X+b2X2+b3X34

3.5對回歸方程進行線性檢驗

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

計算統計量t，計算公式為：t=bi/SEi

其中，SEi為每個非線性系數的斜率標準誤，計算公式為：

其中，Y為回歸方程預測值，與為測定均值。",,,,,,,,,

,,,,,,,,,,

回歸分析結果,,,樣品數：,11,測定次數,4,,,

回歸方程,方程因子,方程因子值,SEi,t-test(t=bi/Sei）,標準估計誤差Sy.x,自由度df=L?R-Rdf,最適方程,結果分析,

"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，無統計學意義,

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，無統計學意義,

,,,,,,,,,,

"臨床標準的線性檢驗,如回歸分析滿足要求，則可省略",,,,,,,,,

如ADL＜5%，則擬合的多項式無臨床意義,,,,,,,,,

樣本號,測量均值,一次方程預期值,二次方程預期值,三次方程預期值,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%,

從上表數據判定絕對偏倚和相對偏倚是否滿足要求（法定或自定義標準）。,,,,,,,,,

可報告區間建立（本標準僅針對上限，下限為定量限）,,,,,,,,,

高值樣本號,理論濃度,樣本稀釋比例,重復測量結果1,重復測量結果2,重復測量結果3,實測均值,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定偏倚為判斷標準，選取三份樣本的相對偏倚均不大于設定偏倚的最大稀釋倍數為方法推薦的最大稀釋倍數（小于此倍數的三份樣本所有稀釋比例的相對偏倚均不大于設定偏倚），測量區間上限與最大稀釋倍數的乘積為該方法可報告區間的上限。,,,,,,,,,

假定10%為偏倚上限，則3份樣本在稀釋倍數100倍及以下時，實測均值與理論值的偏倚均小于±10%，而200倍時兩份樣本偏倚已超過允許偏倚，故本項目最大稀釋倍數為100。,,,,,,,,,

ADL計算：,,,,,,,,,

內容,,,二次方程,三次方程,,,,,

指導原則方法ADL=,,,0.008%,0.006%,,,,,

行標方法ADL中間計算值,,,0.0006,0.0009,,,,,

行標方法ADL=,,,0.003%,0.004%,,,,,

非線性臨界值PctBnd=,,,5%,5%,,,,,

行標公式,,,,指導原則公式,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,,,,,,

,,,,,p(x):最優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,,,,

對數據進行精密度檢驗，線性評價前提,,,,,L：樣本數,,,,

,,,,,ˉc=（y1+y2+y3+……+yn）/n,,,,

指導原則公式,,,行標公式,,n：樣本數乘以重復次數,,,,

,,,,,d:最優擬合方程的階,,,,

,,,,,σ：最優擬合發能給出的回歸標準誤,,,,

,,,,,Rdf:回歸自由度,,,,

,,,,,R：重復測量的次數,,,,

,,,,,C：常數,,,,

,,,,,,,,,,

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計算中間值,CVr計算中間值,,,,

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,,