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Kit for detecting clostridium difficile and application thereof

A detection kit, a technology for Clostridium difficile, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of poor detection effect and high false positive rate, achieve efficient and rapid detection, improve sensitivity, and have good application prospects. Effect

Active Publication Date: 2017-06-09
嘉兴实践医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection sensitivity of PCR gene detection method is too high, but the false positive rate is also high
The two-step enzyme-linked immunoassay also has the problem of poor detection effect, which needs further improvement

Method used

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  • Kit for detecting clostridium difficile and application thereof
  • Kit for detecting clostridium difficile and application thereof
  • Kit for detecting clostridium difficile and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 The present invention is used for detecting the kit of Clostridium difficile

[0025] 1, the composition of kit of the present invention

[0026] (1) diluent of the present invention:

[0027] Formula 1: PBS (PH7.4), 1% NP-40, 0.5% TritonX-100, 1% BSA, 0.05% Tween20

[0028] Formula 2: PBS (PH7.4), 1% NP-40, 1% TritonX-100, 1% BSA, 0.05% Tween20

[0029] Formula 3: PBS (PH7.4), 1% NP-40, 2% TritonX-100, 1% BSA, 0.05% Tween20

[0030] PBS: Phosphate Buffered Saline

[0031] (2) blocking solution of the present invention

[0032] Formula 1: PBS, 2% BSA, 1% trehalose, 1% FBS

[0033] Formula 2: PBS, 2% BSA, 2% trehalose, 2% FBS

[0034] Formula 3: PBS, 2% BSA, 3% trehalose, 3% FBS

[0035] Formula 4: PBS, 2% BSA, 5% trehalose, 5% FBS

[0036] (3) Other components: ELISA plate coated with specific antibody, HRP enzyme-labeled secondary antibody, positive control substance (GLDH recombinant antigen, ToxinA / B recombinant antigen), washing solution, chromo...

experiment example 1

[0044] Comparison of Experimental Example 1 and Existing Universal Diluent

[0045] 1. Experimental materials

[0046] The kit of the present invention: the kit of Example 1 of the present invention is used, wherein the blocking solution is Formula 2, and the diluent of Formula 1 to Formula 10 is used as the diluent.

[0047] Formula 1: PBS (PH7.4), 1% NP-40, 0.5% TritonX-100, 1% BSA, 0.05% Tween20

[0048] Formula 2: PBS (PH7.4), 1% NP-40, 1% TritonX-100, 1% BSA, 0.05% Tween20

[0049] Formula 3: PBS (PH7.4), 1% NP-40, 2% TritonX-100, 1% BSA, 0.05% Tween20

[0050] Formula 4: PBS (PH7.4), 0.1% NP-40, 1% BSA, 0.05% Tween20

[0051]Formula 5: PBS (PH7.4), 1% NP-40, 1% BSA, 0.05% Tween20

[0052] Formula 6: PBS (PH7.4), 5% NP-40, 1% SDS, 1% BSA, 0.05% Tween20

[0053] Formula 7: PBS (PH7.4), 1% NP-40, 1% SDS, 1% BSA, 0.05% Tween20

[0054] Formula 8: PBS (PH7.4), 1% NP-40, 2% SDS, 1% BSA, 0.05% Tween20

[0055] Formula 9: PBS (PH7.4), 1% NP-40, 1% SDS, 0.5% TritonX-100, 1...

experiment example 2

[0072] Comparison of Experimental Example 2 and the Existing Universal Blocking Solution

[0073] 1. Experimental materials

[0074] The kit of the present invention: the kit of Example 1 of the present invention is used, wherein the blocking liquid is formula 1 to formula 4, and the diluent is formula 1 respectively.

[0075] Control kit: except that the diluent is a universal blocking solution, other components are the same as the kit of the present invention, and the universal blocking solution is: PBS (PH7.4), 1% BSA, 0.05% Tween20.

[0076] 2. Experimental method

[0077] Get GLDH recombinant antigen, positive sample 1,2 (toxigenic Clostridium difficile culture supernatant), negative sample (common E. detection.

[0078] The detection method is as follows:

[0079] Use different blocking solutions to block the ELISA plates coated with specific antibodies. After the blocking solutions of each formula are sealed, part of them is stored at 4°C, and the other part is subj...

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PUM

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Abstract

The invention discloses an enzyme-linked immunosorbent assay (ELISA) detection kit for clostridium difficile. The kit comprises the following components: a diluent, confining liquid, a clostridium difficile antibody coated ELISA plate, an ELISA second antibody, a positive reference substance, washing liquid, color developing liquid and stop liquid, wherein every 100ml of diluent contains 1ml of NP-40, 0.5-2ml of Triton X-100, 1g of BSA and 0.05ml of Tween20 phosphate buffer. The invention also discloses an application of the kit in clostridium difficile detection. The kit disclosed by the invention can efficiently detect clostridium difficile and has good application prospects.

Description

technical field [0001] The invention relates to a kit for detecting Clostridium difficile and its application. Background technique [0002] Clostridium difficile (C.DIFF) is a Gram-positive, spore-forming, obligate anaerobic Clostridium, the only anaerobic bacterium that can cause nosocomial infections, and the only pathogen that can cause nosocomial infections that can form spores bacteria. The main cause of diarrhea caused by Clostridium difficile infection is that the use of antibiotics leads to changes in the bacterial composition of the flora, causing dysbiosis, which leads to the proliferation of Clostridium difficile flora, and finally causes diarrhea. Higher rates have been thought to occur in immunocompromised elderly and pediatric patients with prolonged exposure to antibiotics and serious underlying comorbidities. Symptoms of infection can vary widely, from an asymptomatic state or mild C. difficile infection to a severe and life-threatening state. Since the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/56911G01N33/577G01N2333/32
Inventor 黄绣川邱兵
Owner 嘉兴实践医学科技有限公司
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