RNA was isolated from cervical cell lines or cytology samples stored in SurePath™ preservative at room temperature for 2–5 weeks using five commercially available RNA purification kits, three of which contain proteinases. The quality of the RNA was assessed by real time RT-PCR amplification of GAPDH, GUSB, U1A, HPV 16 and 18 E6 mRNAs.
RNA was isolated successfully from cells that were stored in SurePath™ preservative fluid with only the three protocols that contained proteinases. GAPDH was amplified in 98–100 % of the samples, GUSB in 90–98 % , and the least abundant transcript, U1A, was amplified in 81–96 % of the samples. HPV 16 and 18 E6 transcripts were detected in 56 % of high grade, 39 % of low grade and 2 % of normal samples, with a concordance between DNA genotype and E6 mRNA expression of 97 % .
We demonstrated that RNA can be extracted from cervical cell lines and cytology specimens stored in BD SurePath™ preservative fluid with three different procedures that all contain proteinases. This RNA is suitable for real-time RT-PCR applications.