PURPOSE: Detection of variant specific genes by reverse transcription polymerase chain reaction (RT-PCR) has become a routine diagnostic test for accurate subtyping of RNA viruses. The Influenza A (H1N1) virus RNA stability according to storage conditions and duration of swabbed samples and extracted RNA samples are important in research for other respiratory virus and bacterial infection. We have studied Influenza A (H1N1) virus RNA using RT-PCR for the stability in extracted RNA samples and swabbed samples.
METHODS: We have tested 15 Influenza A (H1N1) virus RNA samples and 15 swabbed samples confirmed initially positive by real time RT-PCR. We have studied RNA stability after storage (within 12hours, 4 months and 6 months) at different temperature (-20 and -80 degrees C). Swabbed samples and extracted RNA samples containing different Influenza A (H1N1) virus were stored and analysed by real time RT-PCR at defined time points.
RESULTS: Influenza A (H1N1) virus RNA levels remained almost stable (above 90%) at 4 months and 6 months at -20 and -80 degrees C. However swabbed samples at -20 and -80 degrees C after storage were unstable. We could not detect Influenza A (H1N1) virus in swabbed samples of 3 cases and 5 cases at each temperature.
CONCLUSIONS: We conclude that extracted RNA samples can be stored at -20 degrees C for 6months without any significant difference in detected Influenza A (H1N1) virus RNA by using real time RT-PCR and swabbed sample is suitable for long term storage.
CLINICAL IMPLICATIONS: Results of our study can be used for storage of Influenza virus
DISCLOSURE: The following authors have nothing to disclose: Chang Youl Lee, In-Bum Suh
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