CHARACTERIZATION OF RNA IN CYTOLOGIC SAMPLES PRESERVED IN A METHANOL-BASED COLLECTION SOLUTION

Background: ThinPrep is a fluid-based technique for collection and pro cessing of cytologic specimens. The present study was designed to dete rmine whether the collection solution preserved RNA for molecular anal ysis. Methods and Results: Cervical cancer cell lines and cord blood l ymphocytes were used to test the efficacy of various protocols for fix ation, storage, and extraction of RNA. Total RNA was extracted and ana lyzed by denaturing gel electrophoresis. Preserved cells stored for 24 hours at room temperature or 4 degrees C had intact 28S and 18S ribos omal RNA. Both cellular and viral messenger RNAs were amplified from p reserved samples by reverse transcription polymerase chain reaction (R T-PCR). Viral messenger RNA (mRNA) could be detected in a mixture of p re served cells containing 10% human papillomavirus (HPV) positive cel ls. RNA preservation in clinical samples was adequate for RT-PCR of ce llular mRNA. Conclusions: Both experimental samples and clinical sampl es collected in the preservation media had intact total RNA. Amplifica tion of both cellular and HPV mRNA was successful.