Journal of Plant Pathology (2012) 94, 681-685

W. Sledz, A. Adamowska, J. Piosik and E. Lojkowska (2012)
Detecting live and dead cells of Pectobacterium atrosepticum based on immunomagnetics separation and staining
Journal of Plant Pathology 94 (3), 681-685
Abstract: An assay that combines immunomagnetic separation (IMS) and staining by the LIVE/DEAD BacLight™ Bacterial Viability Kit was developed for the rapid detection and identification of living and dead cells of the plantpathogenic bacterium Pectobacterium atrosepticum (Pba) in potato tuber tissue. In the development of this IMS/LD assay, bacterial suspensions in buffer or potato peel extract were spiked with living and dead cells of Pba and incubated with paramagnetic beads coated with Pbaspecific antibodies. Beads with Pba cells were recovered from the suspensions with a magnet and the immobilized cells were stained with a mixture of Syto9 and propidium iodide. The collected cells were analysed by fluorescence microscopy. The assay required minimal sample preparation (e.g. no wash steps were required) and could be completed within 1 h. The detection limit was 10⁴ and 10⁵ cells per ml for M-280 and M-450 paramagnetic beads, respectively, which is similar to that previously reported for ELISA. The specificity and sensitivity of the IMS/LD assay were similar regardless of whether Pba cells were in potato peel extracts or in buffer. The assay also allowed the differentiation of live and dead Pba cells, which is important in predicting possible losses caused by Pba to potato crops.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Full text of article
Database assignments for author(s): Ewa Lojkowska

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

---

Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Pectobacterium atrosepticum	Potato (Solanum tuberosum)