Preliminary investigation into the use of a real-time PCR method for the quantification of an oligonucleotide in human plasma and the development of novel acceptance criteria.

BACKGROUND: The aim of the work was to evaluate the sensitivity and reproducibility of real-time reverse transcriptase PCR for quantitative analysis of an oligonucleotide in a biological matrix. A novel approach for the identification of outliers when assessing the suitability of calibration standards and QC samples is investigated.
RESULTS: A suitable assay was established for the determination of the oligonucleotide in human plasma over a range of 0.5-100 ng/ml.
CONCLUSION: In these preliminary investigations, the precision and accuracy of the method was established for the quantification of the oligonucleotide in human plasma. It was established that the method was precise and accurate for quantification of the oligonucleotide in human plasma. The acceptability of the data was assessed using a novel three-step process to identify any outliers, involving the use of the Grubbs' test. The analytical method only requires a small sample volume (<0.01 ml), so would be applicable in analysis of low-volume samples.