Visual detection technique for efficient screening and isolation of Salmonella based on a novel enrichment assay using chromatography membrane
To detect Salmonella more efficiently and isolate strains more easily, a novel and simple detection method that uses an enrichment assay and two chromogenic reactions on a chromatography membrane was developed. Grade 3 chromatography paper is used as functionalized solid phase support (SPS), which c...
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Veröffentlicht in: | European journal of clinical microbiology & infectious diseases 2016-03, Vol.35 (3), p.353-361 |
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Hauptverfasser: | , , , , , , , , , , , , , , , , , , , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | To detect
Salmonella
more efficiently and isolate strains more easily, a novel and simple detection method that uses an enrichment assay and two chromogenic reactions on a chromatography membrane was developed. Grade 3 chromatography paper is used as functionalized solid phase support (SPS), which contains specially optimized medium. One reaction for screening is based on the sulfate-reducing capacity of
Salmonella
. Hydrogen sulfide (H
2
S) generated by
Salmonella
reacts with ammonium ferric citrate to produce black colored ferrous sulfide. Another reaction is based on
Salmonella
C8 esterase that is unique for
Enterobacteriaceae
except
Serratia
and interacts with 4-methylumbelliferyl caprylate (MUCAP) to produce fluorescent umbelliferone, which is visible under ultraviolet light. A very low detection limit (10
1
CFU ml
−1
) for
Salmonella
was achieved on the background of 10
5
CFU ml
−1
Escherichia coli
. More importantly, testing with more than 1,000 anal samples indicated that our method has a high positive detection rate and is relatively low cost, compared with the traditional culture-based method. It took only 1 day for the preliminary screening and 2 days to efficiently isolate the
Salmonella
cells, indicating that the new assay is specific, rapid, and simple for
Salmonella
detection. In contrast to the traditional culture-based method, this method can be easily used to screen and isolate targeted strains with the naked eye. The results of quantitative and comparative experiments showed that the visual detection technique is an efficient alternative method for the screening of
Salmonella spp.
in many applications of large-sized samples related to public health surveillance. |
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ISSN: | 0934-9723 1435-4373 |
DOI: | 10.1007/s10096-015-2543-2 |