Interferograms plotted with reference change value (RCV) may facilitate the management of hemolyzed samples

J Med Biochem. 2022 Feb 2;41(1):53-61. doi: 10.5937/jomb0-31250.

Abstract

Background: The European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Preanalytical Phase (WG-PRE) have recommended an algorithm based on the reference change value (RCV) to evaluate hemolysis. We utilized this algorithm to analyze hemolysis-sensitive parameters.

Methods: Two tubes of blood were collected from each of the 10 participants, one of which was subjected to mechanical trauma while the other was centrifuged directly. Subsequently, the samples were diluted with the participant's hemolyzed sample to obtain the desired hemoglobin concentrations (0, 1, 2, 4, 6, 8, and 10 g/L). ALT, AST, K, LDH, T. Bil tests were performed using Beckman Coulter AU680 analyzer. The analytical and clinical cut-offs were based on the biological variation for the allowable imprecision and RCV. The algorithms could report the values directly below the analytical cut-off or those between the analytical and clinical cut-offs with comments. If the change was above the clinical cut-off, the test was rejected. The linear regression was used for interferograms, and the hemoglobin concentrations corresponding to cut-offs were calculated via the interferograms.

Results: The RCV was calculated as 29.6% for ALT. Therefore, ALT should be rejected in samples containing >5.9 g/L hemoglobin. The RCVs for AST, K, LDH, and T. Bil were calculated as 27.9%, 12.1%, 19.2%, and 61.2%, while the samples' hemoglobin concentrations for test rejection were 0.8, 1.6, 0.5, and 2.2 g/L, respectively.

Conclusions: Algorithms prepared with RCV could provide evidence-based results and objectively manage hemolyzed samples.

Uvod: Radna grupa za preanalitičku fazu (WG-PRE) Evropske Federacije za kliničku hemiju i laboratorijsku medicinu (EFLM) preporučila je algoritam koji se zasniva na promeni referentne vrednosti (RCV) za procenu hemolize. Mi smo koristili ovaj algoritam za analiziranje osetljivih hemolizovanih parametara.

Metode: Uzete su dve epruvete krvi od svakog od 10 učesnika, jedna koja je bila predmet mehaničke traume a druga je direktno centrifugirana. Istovremeno uzorci su bili razblaženi sa hemolizovanim uzorcima učesnika da bi se dobile željene koncentracije hemoglobina (0, 1, 2, 4, 6, 8 i 190 g/L). Analiziranje ALT, AST, K, LDH, T. Bil je izve deno primenom Beckman Coulter AU680 analizatora. Analitičke i kliničke cut-off vrednosti bile su zasnovane na biološkoj varijaciji za dozvoljivu nepreciznost i RCV. Algo ritmi su odgovarali direktno vrednostima ispod kliničke cut-off ili one između analitičke i kliničke cut-off. Ako je pro mena bila iznad kliničke cut-off test je odbačen. Linearna regresija je korišćena na interferograme i koncentracije hemo globina koje su odgovarale cut-off vednostima izračunatih preko interferograma.

Rezultati: RCV je izračunat kao 29,6% za ALT. Prema tome potrebno je odbaciti uzorke za ALT koji su imali >5,9 g/L hemoglobina. RCV za AST, K, LDH i T. Bil izračunati su kao 27,9%, 12,1%, 19,2% i 61,2%, dok su uzorci sa koncentracijom hemoglobina za odbacivanje testa bili 0,8, 1,6, 0,5 i 2,2 g/L.

Zaključak: Algoritimi pripremljeni sa RCV mogu da obezbe rezultate zasnovane na dokazima i objektivno procene hemolizovane uzorke.

Keywords: hemolysis; interference; laboratory errors; preanalytical phase; reference change value.