Food poisoning by Diarrheagenic Escherichia coli is an infectious food poisoning that is caused by the growth of E.coli taken into the intestinal tract though the ingestion of food. These Diarrheagenic E.coli can be broadly classified into five types.1) Among these, Enterohemorrhagic E.coli (EHEC) causes hemorrhagic colitis through the action of the verotoxin that it produces; it can also cause hemolytic uremic syndrome and encephalopathy, which can be fatal. Many cases of mass food poisoning by EHEC have occurred, so EHEC is viewed as the most dangerous among Diarrheagenic E.coli from the standpoint of food hygiene. The diarrheagenic E.coli, O111, has been detected as the second commonest serotype after O157 in foreign countries 2) and reported to be the third commonest serotype after O157 and O26 in Japan 3).
This product is a kit for detecting E.coli O111 in foods by immunochromatography. Tests can be conducted rapidly and simply by means of the kit.
Detection of E.coli O111 in Foods
Note 1: This kit is intended to specifically detect E.coli O111 and therefore cannot detect the presence or absence of verotoxin prodution
(1) The simple one-step operation of the kit.
(2) The test gives rapid results.
(3) There is no need for special test equipment.
A:Test plate 5-test 4 packs
B:Instruction manual 1 sheet
2) Ingredients and Quantity
A:Anti-E.coli O111 polyclonal antibody (rabbit) 0.375 ug
B:Anti-E.coli O111 polyclonal antibody labeled with colloidal gold (rabbit) 0.13 ug
C:Anti-rabbit immunoglobulin polyclonal antibody (goat) 0.25 ug
1) Illustration of Test plate
a. Sample solution drop section (Be careful not to touch this sectoin with your finger.)
b. Reagent-containing section
c. Detecting section (Be careful not to scratch this section and touch this section with your finger.)
d. Absorbent pad
e. Measurement items listing position
f. Test line appearance position (Approx.30mm from the sample solution drop section.)
g. Control line appearance position (Approx.38mm from the sample solution drop section)
2) Principle of assay
When a sample solution is dropped onto the test sample drop section of the test plate, the gold colloid-labeled anti-E. coli O111 polycolonal antibody (2) in the test sample-containing section dissolves and forms complexes with E. coli O111 (1). These complexes move to the expanded section by capillary attraction and are trapped by the anti-E. coli O111 antibody (3) that is fixed in the test line appearance position. This results in the appearance of a reddish purple line of gold colloid. This reddish purple line can be detected by visual inspection and used to judge the presence or absence of E. coli O111 in the test solution. The excess gold-labeled antibodies, regardless of the presence or absence of E. coli O111 in the test solution, travel further through the expanded section and are trapped by the anti-goat immunoglobulin rabbit antibody(4) fixed at the control line appearance position, where they form a second reddish purple line. The presence of this line indicates that the test solution has reached the expanded section.
The method for preparing a sample solution is described in accordance with the Japanese official testing method More
1) NH Immunochromato O111 Test Procedures
(1) Bring the test plates contained in the aluminum pouch to room temperature and remove from the pouch as necessary immediately before use.
(2) With an oil-based marker pen, write the name of the test sample or the number of the subject under test on the absorbent pad of the test plate removed from the bag.
(3) Place the test plate carefully on the flat stand and drop a 100 μL-portion of the test solution onto the test sample drop section (see the figure on the left). Otherwise, dispense a 150 μL portion of the test solution into a test tube and attach the test plate to the test tube so that the test sample drop section of the test plate is immersed in the test solution (see the figure on the right).
(4) Allow the test plate to stand undisturbed for 15 minutes and then visually judge the presence or absence of E. coli O111 in the solution.
Note1: Do not remove the test plate from the aluminum pouch until it has returned to room temperature, otherwise incorrect test results may be obtained as a result of moisture absorption.
Note2: Keep test plates that are not in use in a vinyl pouch containing desiccants and store this in an aluminum pouch in a refrigerator.
Note3: Be careful not to scratch the test sample drop section or expanded section and do not touch them with your hands. When handling the test plate, make sure that you hold the absorbent pad.
Note4: Make sure that you use a sterilized pipette or chip to drop or dispense the sample solution. Change the pipette or chip for every test solution.
Note5: Make sure that the 150-μL portion of test solution does not overflow the test plate when dropping it. If necessary, drop the solution in two or more portions.
Note6: To prevent infection of the operator, it is recommended that testing be performed with the test plate. A wrap should be placed under the plate. when dropping the test solution.
2) Judgment of the Test Results
(1) The test results is judged as positive when a reddish purple line is observed at the test line appearance position and at the control line appearance position 15 minutes after the start of the test.*
(2) Judge the test results as negative when no reddish purple line is observed at the test line appearance position, but a line is observed at the control line appearance position.
(3) Retest in cases where no reddish purple line is observed at the control appearance position, regardless of the presence or absence of a line at the test line appearance position. It is likely that there is something abnormal in the development of the sample solution in such cases.
Note1: Ensure that confirmatory tests, in accordance with the official test method or other methods, are performed on specimens that are tested positive by the kit. The proliferated and cultivated samples used for testing with the kit can be used in confirmatory tests by the official or other test method.
NH Immunochromato O111
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.