An optimised for the extraction of non-viral mRNA from human plasma frozen for three years

Sze Chuen Cesar Wong, E. S F Lo, M. T. Cheung

Research output: Journal article publicationJournal articleAcademic researchpeer-review

16 Citations (Scopus)

Abstract

Aims: To detect non-viral mRNA in human plasma that has been frozen for three years using a new protocol. Methods: Plasma from 15 patients with colorectal cancer and 10 normal subjects was separated and frozen with Trizol at -80°C for three years. As a control measure, plasma from 10 of the 15 patients was separated using the same protocol but no Trizol during storage. After three years, all samples were extracted using Trizol and RNeasy before the reverse transcriptose polymerase chain reaction was performed to detect non-viral β catenin mRNA. In addition, extraction of three plasma samples by Trizol or RNeasy independently was carried out for comparison. Results: β Catenin mRNA was detected in all 15 patient plasma samples and only one of the 10 normal subjects. In contrast, no β catenin mRNA was found in the control and patient samples that were independently extracted by Trizol and RNeasy kit. Conclusions: This new protocol is a reliable method for extracting non-viral mRNA from the plasma of patients with cancer after longterm storage for three years. Extractions using Trizol and RNeasy kits independently could not isolate mRNA with sufficient quantity and quality for detection.
Original languageEnglish
Pages (from-to)766-768
Number of pages3
JournalJournal of Clinical Pathology
Volume57
Issue number7
DOIs
Publication statusPublished - 1 Jul 2004
Externally publishedYes

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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