Sepsis pp 65-72 | Cite as

Broad-Range PCR in the Identification of Bacterial and Fungal Pathogens from Positive Blood Culture Bottles: A Sequencing Approach

  • Yoshitomo Morinaga
  • Katsunori YanagiharaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1237)


Rapid identification of causative bacteria in patients with sepsis can contribute to appropriate selection of antibiotics and improvement of patients’ prognosis. Genotypic identification is an emerging technology that may provide an alternative method to, or complement, established phenotypic identification procedures.

Sequence analysis of the 16S rRNA gene is a widely accepted tool for molecular identification of bacteria. Pyrosequencing is a DNA sequencing technique that is based on the detection of pyrophosphate that is released during DNA synthesis. Pyrosequencing can provide sequence information rapidly by reading short sequences; therefore, it may contribute to a rapid identification and lead to a great help in improving the outcome of sepsis. The DNA pyrosequencing-based identification from positive blood culture samples basically consisted of the following four steps: (1) DNA extraction, (2) amplification of target genes, (3) DNA pyrosequencing, and (4) homology searching.

Key words

Pyrosequence Genetic identification 16S rRNA V1 V3 Sepsis 



We thank Maiko Motoshima and Sayaka Mori for technical assistance.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Laboratory MedicineNagasaki University Graduate School of Biomedical SciencesNagasakiJapan

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