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Validation of an enzyme-linked immunosorbent assay for C-peptide analysis in Cameroon
M Asanghanwa1, F van Genderen, K Verhaeghen
1Diabetes Research Centre, Brussels Free University, B-1090 Brussels, Belgium.
Aims:
To validate an ELISA method for C-peptide analysis in Cameroon.
Methods:
We evaluated the linearity, detection limit, functional sensitivity, precision and accuracy, and further investigated for cross-reactivity by proinsulin, and interferences by lipids, bilirubin and hemoglobin. This method was compared with the Roche electrochemiluminescence immunoassay. C-peptide stability was assessed following a series of freeze-thaw cycles, and after storage at room temperature. The C-peptide reference range was determined by analyzing fifty plasma samples of Cameroonians without diabetes.
Results:
The ELISA was linear at least up to 7.09 μg/L, and had a detection limit of 0.09 μg/L, and a functional sensitivity of 0.32 μg/L. The inter- and intraassay %CV were 2.9-9.9%, and 5.2-9.4%, respectively. Recoveries were 81-94% in serum, and 93-98% in buffer. Comparison with the ECLIA yielded a good correlation coefficient (R(2)=0.98). There was no cross-reactivity with proinsulin, and no interference with lipids, bilirubin and hemoglobin. C-peptide was stable at room temperature for 24 h and up to 7 freeze-thaw cycles for medium (1-6 μg/L) and high (>6 μg/L) levels (<-15°C and <-70°C). The reference range for C-peptide was 0.38-3.63 μg/L.
Conclusions:
This method is suitable for C-peptide analysis in low-income countries like Cameroon.
