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Automated high throughput E. coli o157:H7 PCR detection system and uses thereof
Automated high throughput E. coli o157:H7 PCR detection system and uses thereof
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机译：自动化的高通量大肠杆菌o157：H7 PCR检测系统及其用途
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摘要
The present invention describes the development and evaluation of the sensitivity and specificity of a PCR-based 5′ nuclease assay for presumptively detecting E. coli O157:H7 DNA. The specificity of the eaeA-based, 5′ nuclease assay system was sufficient to correctly identify all E. coli O157:H7 strains evaluated, mirroring the previously described specificity of the PCR primers. The SZ-primed, eaeA-targeted, 5′ nuclease detection assay was capable of rapid, semiautomated, presumptive detection of E. coli O157:H7 when ≧103 CFU/ml were present in modified tryptic soy broth (mTSB) or modified E. coli broth (mEC), and when ≧104 CFU/ml were present in ground beef-mTSB mixtures. Incorporating an immunomagnetic separation step (IMS), followed by a secondary enrichment culturing step and with DNA recovery, improved the detection threshold to ≧102 CFU/ml.
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