Effects of Different Lysis Buffers of Nucleic Acid Purification Kit on the Stability of Influenza Virus RNA

ABSTRACT 

Aim: Under suboptimal storage and transport conditions, influenza virus (flu-v) RNA is prone to degradation and lysis buffers from RNA extraction kits have a potential to stabilize RNA. The aim of this study was to investigate the effects of different lysis buffers on the stability of flu-v RNA. Materials & methods: Aliquots of flu-v suspension were processed in parallel with two lysis buffers, and then underwent cyclic freeze–thaw or prolonged storage at 4, 22 and -20°C. The viral RNA was analyzed by using real-time and conventional RT-PCR amplifying, respectively, partial and full-length sequences of the flu-v matrix gene. Results: The viral RNA remained intact in samples treated with either of the two lysis buffers for at least 7 days at 4°C, 90 days at -20°C or following seven freeze–thaw cycles, but buffer A was superior to buffer B in protecting RNA from degradation at 4°C and 22°C, or following a further increase of freeze–thaw cycles. Conclusion: Lysis buffer preservatives provide viral RNA stabilization, whereas different lysis buffers vary in their ability to stabilize viral RNA, and thus their performance characteristics should be evaluated prior to their application in clinical practice.

KEYWORDS: :

• biological preservation
• buffer
• freezing
• guanidine
• influenza A virus
• reverse transcriptase polymerase chain reaction
• RNA stability
• specimen handling
• temperature
• time factors
• viral RNA

View correction statement:

Erratum

Financial & competing interests disclosure

This research was supported by the State Project on Major Infectious Diseases Prevention (2012ZX10002006-003) and Program for Changjiang Scholars and Innovative Research Team in University (grant no. IRT1131). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Additional information

Funding

This research was supported by the State Project on Major Infectious Diseases Prevention (2012ZX10002006-003) and Program for Changjiang Scholars and Innovative Research Team in University (grant no. IRT1131). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.