Pre-analytical stability of serum biomarkers for neurological disease: neurofilament-light, glial fibrillary acidic protein and contactin-1.

Zoë Y G J van Lierop, Inge M W Verberk, Kees W J van Uffelen, Marleen J A Koel-Simmelink, Lisanne In 't Veld, Joep Killestein, Charlotte E Teunissen

Clinical chemistry and laboratory medicine 2022 May 25

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Neurofilament-light (NfL), glial fibrillary acidic protein (GFAP) and contactin-1 (CNTN1) are blood-based biomarkers that could contribute to monitoring and prediction of disease and treatment outcomes in neurological diseases. Pre-analytical sample handling might affect results, which could be disease-dependent. We tested common handling variations in serum of volunteers as well as in a defined group of patients with multiple sclerosis (pwMS). Sample sets from 5 pwMS and 5 volunteers at the outpatient clinic were collected per experiment. We investigated the effect of the following variables: collection tube type, delayed centrifugation, centrifugation temperature, delayed storage after centrifugation and freeze-thawing. NfL and GFAP were measured by Simoa and CNTN1 by Luminex. A median recovery of 90-110% was considered stable. For most pre-analytical variables, serum NfL and CNTN1 levels remained unaffected. In the total group, NfL levels increased (121%) after 6 h of delay at 2-8 °C until centrifugation, while no significant changes were observed after 24 h delay at room temperature (RT). In pwMS specifically, CNTN1 levels increased from additional freeze-thaw cycles number 2 to 4 (111%-141%), whereas volunteer levels remained stable. GFAP showed good stability for all pre-analytical variables. Overall, the serum biomarkers tested were relatively unaffected by variations in sample handling. For serum NfL, we recommend storage at RT before centrifugation at 2-8 °C up to 6 h or at RT up to 24 h. For serum CNTN1, we advise a maximum of two freeze-thaw cycles. Our results confirm and expand on recently launched consensus standardized operating procedures. © 2022 Zoë Y. G. J. van Lierop et al., published by De Gruyter, Berlin/Boston.

Citation

Zoë Y G J van Lierop, Inge M W Verberk, Kees W J van Uffelen, Marleen J A Koel-Simmelink, Lisanne In 't Veld, Joep Killestein, Charlotte E Teunissen. Pre-analytical stability of serum biomarkers for neurological disease: neurofilament-light, glial fibrillary acidic protein and contactin-1. Clinical chemistry and laboratory medicine. 2022 May 25;60(6):842-850