E.coli O157:H7 Pincer^TM Assay

Catalog No: EC1014

Escherichia coli (E. coli) is a Gram negative bacterium that commonly lives in the intestines of people and animals. Most of the E. coli strains are normal inhabitants of the small intestine and colon and are non-pathogenic. Pathogenic E.coli strains have been classified into three classes: ETEC (enterotoxigenic), EPEC (enteropathogenic) and EHEC (enterohemorrhagic). E.coli serotype O157:H7 is a rare variety of E.coli strain that belongs to EHEC. It causes colitis and bloody diarrhea by producing a toxin called Shiga toxin, which damages the intestines. This toxin is closely related to or identical to the toxin produced by Shigella dysenteriae. Serotype O157:H7 is estimated to cause infection in more than 70,000 patients a year in the United States. It has been reported to cause both large outbreaks as well as isolated sporadic infections in small numbers of individuals (http://ecoliinformation.com).

Typical isolation and identification methods for O157:H7 are time consuming and easily interfered by background microorganisms. The E.coli O157:H7 PincerTM kit is designed for rapid qualitative and quantitative assessment of E.coli O157:H7 in pre-enriched samples.

This assay is a rapid, homogeneous antibody-based fluorescence assay for detecting E.coli O157:H7 in food, water or other environmental samples. The assay uses two E.coli O157:H7 pincers that are anti-E.coli (serotype O157:H7) specific antibody conjugated to short complementary single-stranded oligonucleotides. In the presence of E.coli, the two single-stranded oligos associate to form a duplex and FRET (Fluorescence resonance energy transfer) is initiated between the two fluorophores labelled at the end of the two oligonucleotides¹. This assay is simple (one step) and fast (30 min).

E.coli O157:H7 Pincer^TM Assay - Standard Curves

The following E.coli O157:H7 standard curves were prepared using the E.coli O157:H7 Pincer^TM Assay kit with the fluorescence signal read at 30 minutes after addition of Pincer^TM Assay Solution.

E.coli O157:H7 Pincer^TM Assay – Sensitivity And Specificity

Sensitivity: The minimum detectable bacterial number by this assay is estimated to be 1000 cells/20μl.

Detection range: 10³ - 10⁵ cells/20μl

Specificity: E.coli O157:H7 Pincer^TM Assay Kit is specific to E.coli O157:H7 and will not cross react with various related antigens.

Reference:

1. Heyduk T, & Heyduk, E, (2005) Anal.Chem. 77:1147-1156.

RESEARCH USE:

For research use only, not for use in diagnostic procedures.

Patent pending.

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Product name	Measurements	Cat. #	Protocol	MSDS	Price	Order
E.coli O157:H7 PincerTMAssay kit	100	EC1014-100			$240
E.coli O157:H7 PincerTMAssay kit	384	EC1014-384			$360
E.coli O157:H7 PincerTMAssay kit	384 x 2	EC1014-384 x 2			$680
E.coli O157:H7 PincerTMAssay kit	384 x 3	EC1014-384 x 3			$900
384-well round-bottom low volume non-binding surface black polystyrene microplate	1 plate	163301	--		$9.50