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The quality of the experiment and the estimates of systematic errors will depend more on getting a wide range of test results than a large number of test results.
Duplicate analyses would demonstrate whether or not these observed discrepancies were repeatable.
If duplicates are not performed, then it is critical to inspect the comparison results at the time they are collected, identify those specimens where the differences are large, and repeat those analyses while the specimens are still available.
Stability may be improved for some tests by adding preservatives, separating the serum or plasma from the cells, refrigeration, or freezing.
Specimen handling needs to be carefully defined and systematized prior to beginning the comparison of methods study.
Ideally, these duplicates should be two different samples (or cups) that are analyzed in different runs, or at least in different order (rather than back-to-back replicates on the same cup of sample).
The duplicates provide a check on the validity of the measurements by the individual methods and help identify problems arising from sample mix-ups, transposition errors, and other mistakes.
Several different analytical runs on different days should be included to minimize any systematic errors that might occur in a single run.
A minimum of 5 days is recommended , but it may be preferable to extend the experiment for a longer period of time.
Any differences between a test method and a routine method must be carefully interpreted.
If the differences are small, then the two methods have the same relative accuracy.