Long-term conservation of HCV RNA at 4 degrees C using a new RNA stabilizing solution

Area of Science:

• Molecular Biology
• Biochemistry
• Virology

Background:

• RNA is highly susceptible to degradation by ribonucleases, especially at temperatures above 0°C.
• Maintaining RNA integrity and biological activity is critical for molecular biology applications.
• Existing methods for RNA preservation often require freezing, limiting accessibility and increasing costs.

Purpose of the Study:

• To evaluate the efficacy of a new stabilizing solution for long-term RNA preservation.
• To assess RNA stability at 4°C and -20°C using the novel solution.
• To determine the suitability of the solution for storing both naked RNA and RNA within clinical samples.

Main Methods:

• RNA stability was tested using a stabilizing solution containing chaotropic agents, buffer, phenol, and yeast RNA.
• Two RNA substrates were used: naked RNA (purified or in vitro transcribed) and RNA in hepatitis C virus (HCV)-positive human plasma.
• Stability was assessed by viral load estimation using a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay.

Main Results:

• Naked RNA purified from HCV-positive plasma remained stable at 4°C for at least 24 months.
• An in vitro transcribed RNA standard maintained viability after 36 months of storage at 4°C.
• HCV-positive human plasma dilutions were stable in the solution at 4°C for at least 5 months.

Conclusions:

• The novel stabilizing solution provides long-term naked RNA stability at 4°C.
• This solution is ideal for storing RNA controls and standards for molecular diagnostics.
• The solution can be used for preserving clinical samples before transport to laboratories.