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Simultaneous and highly sensitive detection of six different foodborne pathogens by high-throughput suspension array technology.
- Source :
-
Food Control . Jun2014, Vol. 40, p300-309. 10p. - Publication Year :
- 2014
-
Abstract
- Abstract: In the present paper, multiplex polymerase chain reaction (PCR) was coupled to a suspension array to construct an assay for simultaneous identification of six kinds of pathogens. The suspension array allows the simultaneous detection of different target sequences in a multiplex and high-throughput format. The assay uses a liquid suspension hybridization format with specific oligonucleotide probes covalently bound to the surface of fluorescent color-coded microspheres. Biotinylated target amplicons, hybridized to their complementary probe sequences, are quantified by adding the conjugate, streptavidin R-phycoerythrin. Six probes derived from the sequence analysis of a specific gene were developed and validated. There was little cross-reaction among the various probes. In the multi-channel method, the detection sensitivity was 1.6 × 10−6 mM. Results of nucleic acid detection assays show detection sensitivity for 20–4 × 103 CFU/ml, which is one to five orders of magnitude higher than those of PCR-agarose method in different bacteria. Aside from single channel capability, the assay allows the simultaneous detection of target genes in a single reaction, with detection limits of 1–10 CFU/ml. The accuracy, speed, flexibility, and sensitivity of the proposed assay are beneficial for the diagnosis of pathogenic diseases. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 09567135
- Volume :
- 40
- Database :
- Academic Search Index
- Journal :
- Food Control
- Publication Type :
- Academic Journal
- Accession number :
- 94050174
- Full Text :
- https://doi.org/10.1016/j.foodcont.2013.11.028