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Sensitization test strip for rapidly detecting cronobacter sakazakii and application of sensitization test strip

A technology for test strips and bacilli, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of cumbersome methods, low sensitivity, long time consumption, etc., and achieve the effects of increasing sensitivity, improving sensitivity, and increasing binding amount.

Inactive Publication Date: 2019-04-23
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of low sensitivity of existing traditional test strips and the tedious, complicated and time-consuming methods for improving the sensitivity, and even the need to introduce additional reagents, the present invention provides a new method for rapid detection of Cronobacter sakazakii Sensitization test strips, compared with traditional test strips, this test strip has significantly improved sensitivity, does not require additional reagents, is low in cost, and has a simple preparation method for the test strips, enabling rapid detection of Cronobacter sakazakii The technical scheme adopted for detection is as follows:

Method used

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  • Sensitization test strip for rapidly detecting cronobacter sakazakii and application of sensitization test strip
  • Sensitization test strip for rapidly detecting cronobacter sakazakii and application of sensitization test strip
  • Sensitization test strip for rapidly detecting cronobacter sakazakii and application of sensitization test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Optimization of kit preparation conditions

[0040] 1. Optimization of the concentration of colloidal gold and anti-Kronobacter sakazaki capture antibody in the detection line solution

[0041] 1.1 Preparation of test line solution

[0042] First, the colloidal gold solution is centrifuged and then reconstituted to obtain a concentrated colloidal gold. This makes it easier for the colloidal gold concentration in the incubation system to reach 20PM-80PM. The specific method is as follows: every 1mL colloidal gold solution (with a particle size of 35nm ) Centrifuge at 7000rpm for 20min, remove the supernatant, and then reconstitute it with 10mM Tirs / HCl with a pH of 9 to obtain concentrated colloidal gold. (This concentration step is not limited to this method, and other similar methods can also be used, as long as the concentration of colloidal gold in the detection line solution is 20PM-80PM; and this step can be used or not used in the preparation of the detection...

Embodiment 2

[0056] Example 2: Preparation and application of the sensitization test strip of the present invention

[0057] 1. Preparation of test strips

[0058] This embodiment provides a sensitization test strip for rapid detection of Kronobacter sakazakii. The sensitization test strip includes a sample pad, a gold label pad, an NC film, an absorbent pad, a bottom plate, and a test line on the NC film. And the control line, the control line is coated with a goat anti-mouse secondary antibody; the detection line solution coated on the detection line is obtained by incubating the colloidal gold with the anti-Kronobacter sakazaki capture antibody.

[0059] The preparation method of the detection line solution is as follows: first concentrate the colloidal gold, the specific method is as follows: each 1mL colloidal gold solution (particle size is 35nm) is centrifuged at 7000rpm for 20min, the supernatant is removed, and then the 10mM Tirs / HCl with pH 9 Reconstituted to obtain concentrated colloi...

Embodiment example 3

[0082] Implementation case 3: Comparison of traditional test strips and sensitivity test strips

[0083] This example combines the method of the present invention to prepare a traditional test strip as a control experiment, and compares it with the sensitizing test strip of the present invention. The specific method is as follows:

[0084] 1. Preparation of gold standard solution

[0085] Add potassium carbonate to each 1mL colloidal gold solution to adjust the pH value of the solution, then add 10ug of anti-Kronobacter sakazaki detection antibody, mix well, let stand at room temperature for more than 1h, add 100uL 10% BSA blocking agent, mix well, and stand at room temperature Reaction for more than 1h; after centrifugation, remove the supernatant, add 150uL recombination solution (1% BSA, 2% trehalose, 10mM Tris / HCl) and 2uL 10% BSA precipitation and mix well to obtain the gold standard solution, store the gold standard solution in 4℃ for use.

[0086] 1.2 Assemble traditional test...

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Abstract

The invention discloses a sensitization test strip for rapidly detecting cronobacter sakazakii and application of the sensitization test strip, and belongs to the technical field of detection of cronobacter sakazakii. The sensitization test strip comprises a sample pad, a gold standard pad, an NC film, an absorbent pad and a bottom plate, wherein the NC film is provided with a detection line and acontrol line, the control line is coated with a goat anti-mouse secondary antibody, and a detection line solution coating the detection line is obtained by incubating colloidal gold and an anti-cronobacter sakazakii capture antibody. The sensitization test strip has the characteristic of being capable of improving the protein binding quantity on the NC film, thereby enhancing the capture rate; the colloidal gold added at the detection line can improve aggregation coloration, thereby improving the detection effect; and the sensitivity is improved by 100 times compared with a traditional test strip, and the sensitization test strip has high specificity, short detection time, easy manufacture and a low cost, and is easy to carry and suitable for on-site detection of cronobacter sakazakii.

Description

Technical field [0001] The invention relates to a sensitizing test strip for rapidly detecting Kronobacter sakazakii and its application, and belongs to the technical field of Kronobacter sakazaki detection. Background technique [0002] Kronobacter sakazaki belongs to the genus Kronobacter and is an important food-borne pathogenic bacteria that can cause severe neonatal meningitis, necrotizing enterocolitis and bacteremia. Although the morbidity caused by Kronobacter sakazakii is very low, its fatality rate can reach 80%, and the surviving patients often have serious sequelae of nervous system damage and developmental disorders. Investigations in several disease incidents caused by Sakazaki Krono found that there is a close relationship between Sakazaki Krono infection and infant formula milk powder. Therefore, it is particularly important to establish a rapid and efficient method for the detection of Kronobacter sakazakii in milk powder. [0003] The traditional isolation and d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/532
CPCG01N33/532G01N33/558G01N33/56911
Inventor 潘瑞丽党芳芳王蕊孙露宏付世骞姜毓君满朝新
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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