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Virol J. 2013 Sep 12;10:284. doi: 10.1186/1743-422X-10-284.

High resolution melting analysis: rapid and precise characterisation of recombinant influenza A genomes.

Author information

1
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Südufer 10, Greifswald-Insel Riems, 17493, Germany. donata.kalthoff@fli.bund.de.

Abstract

BACKGROUND:

High resolution melting analysis (HRM) is a rapid and cost-effective technique for the characterisation of PCR amplicons. Because the reverse genetics of segmented influenza A viruses allows the generation of numerous influenza A virus reassortants within a short time, methods for the rapid selection of the correct recombinants are very useful.

METHODS:

PCR primer pairs covering the single nucleotide polymorphism (SNP) positions of two different influenza A H5N1 strains were designed. Reassortants of the two different H5N1 isolates were used as a model to prove the suitability of HRM for the selection of the correct recombinants. Furthermore, two different cycler instruments were compared.

RESULTS:

Both cycler instruments generated comparable average melting peaks, which allowed the easy identification and selection of the correct cloned segments or reassorted viruses.

CONCLUSIONS:

HRM is a highly suitable method for the rapid and precise characterisation of cloned influenza A genomes.

PMID:
24028349
PMCID:
PMC3847122
DOI:
10.1186/1743-422X-10-284
[Indexed for MEDLINE]
Free PMC Article

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