Reference Interval Tutorial CP2308 Clinical Chemistry

Focused reading
Please read pages 251 260: Chapter 14, Establishment and use of reference
values; Burtis CA, Ashwood ER. (eds) Tietz Fundamentals of clinical chemistry 5 th
Edition WB Saunders 2001.

Aim
To determine reference intervals from values in Table 1 using the parametric and
non-parametric statistical methods.

Non-Parametric method
This method is based on rank numbers.
Sort the n reference values in ascending order of magnitude. n is the total
number of reference numbers obtained.
Rank the values. The minimum value has rank number 1; the next value
has rank number 2, and so on until the maximum value which has rank
number n. Consecutive rank numbers should be given to two or more
values that are equal.
Compute the rank number of the 2.5 and 97.5 percentiles is computed as
0.025(n + 1) and 0.975(n + 1) respectively.
One determines the percentile by finding the original reference value that
corresponds to the computed rank numbers, provided the rank numbers
are integers. Otherwise interpolation between the two limiting values is
necessary.

Parametric method
This method assumes that the observed values follow a Gaussian probability
curve. The 2.5 and 97.5 percentiles can be estimated by the values approximately
two SDs on each side of the mean.
Calculate the mean of the given set of data.
Calculate the standard deviation of the given set of data.
Calculate the 2.5 and 97.5 percentile using (x - 1.96SD) and( x +
1.96SD)respectively

Assignment
You should perform your all of your workings in Excel worksheet. Please submit
your worked answer on Blackboard.
1. Non-parametric method analysis: tabulation of rank data; calculation of rank
numbers and reference interval for 2.5 and 97.5 percentile.
2. Parametric method analysis: calculation of mean and SD; calculation of
reference interval for 2.5 and 97.5 percentile.
3. Comment on the reference intervals calculated by both methods
4. Discuss the merits of each method

1
School of Chemical & Life Sciences
Singapore Polytechnic
Reference Interval Tutorial CP2308 Clinical Chemistry

Table 1: Reference values from 120 subjects

Triglyceride Values (mmol/L) from 120 Healthy Individuals

1.
1.73 0.80 1.12 2.50 2.18 1.65
2.
1.68 1.08 1.92 2.55 1.97 1.90
3.
1.61 0.95 1.75 2.62 2.15 1.86
4.
1.72 1.35 0.83 2.10 1.69 2.05
5.
1.76 1.35 2.16 2.65 1.20 1.86
6.
1.64 1.34 1.96 2.72 1.56 1.56
7.
0.51 1.56 1.79 2.80 0.75 1.45
8.
1.57 1.70 1.72 1.49 0.85 1.34
9.
1.45 1.90 1.54 2.91 1.87 1.84
10.
1.65 1.92 1.75 0.43 1.96 1.15
11.
1.78 2.52 1.46 2.26 2.02 1.78
12.
1.95 1.65 1.81 2.27 1.76 1.73
13.
1.10 2.58 1.85 2.27 1.65 1.22
14.
2.30 1.78 2.16 2.28 1.75 1.76
15.
2.35 1.80 2.15 2.30 1.85 1.92
16.
2.48 1.82 1.92 2.31 1.68 2.11
17.
2.51 1.50 1.33 2.34 1.73 2.12
18.
1.25 1.30 1.31 2.35 2.11 0.84
19.
0.65 1.56 2.05 2.48 1.80 1.75
20.
1.71 1.85 1.57 2.21 2.18 1.55

2
School of Chemical & Life Sciences
Singapore Polytechnic