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Escherichia coli kit and identification method

A technology of Escherichia coli and detection reagents, applied in the field of identification of pathogenic bacteria

Active Publication Date: 2021-07-27
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] but previously identified dry chemical enzymatic efficiencies for Escherichia coli or Shigella

Method used

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  • Escherichia coli kit and identification method
  • Escherichia coli kit and identification method
  • Escherichia coli kit and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Plate preparation and interpretation by dry chemical enzymatic method

[0039] Sample bacteria suspension ①: Escherichia coli suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0040] Sample bacterial suspension ②: Shigella sonnei suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0041] Sample bacterial suspension ③: Shigella flexneri suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0042] Sample bacteria suspension ④: Shigella baumannii suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0043] Sample bacterial suspension ⑤: Shigella dysenteriae suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0044] Positive quality control bacteria solution: Escherichia coli suspension, the concentration is 1×10 8 ~5×10 10 individual / mL.

[0045] Sealing agent: liquid paraffin.

[0046] Reaction substrate: 5-bromo-6-chloro-3-indole-β-D galactopyranoside aqueous solution with a con...

Embodiment 2

[0062] Embodiment 2: the influence of bacterial suspension concentration on detection result

[0063] Sample bacteria suspension: prepare different concentrations of Escherichia coli suspension, 1×10 8 pcs / mL, 5×10 8 pcs / mL, 1×10 9 pcs / mL, 5×10 9 pcs / mL, 1×10 10 pcs / mL, 5×10 10 pcs / mL, 7×10 10 individual / mL.

[0064] The preparation method and detection steps of the positive quality control bacteria solution, blocking agent, reaction substrate, chromogenic substrate and plate card are the same as in Example 1;

[0065] Test results:

[0066] The concentration is 1×10 9 pcs / mL, 5×10 9 pcs / mL, 1×10 10 pcs / mL, 5×10 10 The detection of the bacterial suspension of individual / mL is consistent with the detection result of embodiment 1. It shows that in this concentration range, the accurate detection of Escherichia coli can be realized.

[0067] The concentration is 7×10 10 pcs / mL bacterial suspension, after 80 minutes, it was observed that sporadic irregular purplish r...

Embodiment 3

[0068] Embodiment 3: the influence of strain on detection result

[0069] Sample bacterial suspension: prepare bacterial suspensions of different serotypes of Escherichia coli at a concentration of 1×10 8 ~5×10 10 Individuals / mL, including: enteropathogenic Escherichia coli (EPEC), enterotoxigenic Escherichia coli (ETEC), enteroinvasive Escherichia coli (EIEC), enterohemorrhagic Escherichia coli (EHEC ), enteroaggregative Escherichia coli (EAEC).

[0070] The preparation method and detection steps of the positive quality control bacteria solution, blocking agent, reaction substrate, chromogenic substrate and plate card are the same as in Example 1;

[0071] Detection results: the detection results of Escherichia coli suspensions of different serotypes are consistent with the detection results of Example 1. It shows that in this concentration range, the accurate detection of Escherichia coli can be realized.

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Abstract

The invention relates to the technical field of pathogenic bacteria identification, in particular to an Escherichia coli kit and an identification method. The detection reagent comprises a reaction matrix, a reaction substrate and a chromogenic substrate. The reagent has few requirements for sample pretreatment reagent types, the operation complexity is greatly reduced, a sample pretreatment substrate adopts a common chemical reagent, no safety risk exists, and meanwhile the reagent is low in price, easy to obtain and low in cost; besides, the enzyme is adopted to react with the reaction substrate, so that the consumed time is short, and the whole color can be developed, the judgment is simple, and misjudgment is not easy. The reaction result is amaranth in color, the color contrast is obvious, no obvious color difference exists, the result can be directly judged, the operation is simple, the requirement on detection personnel is low, and the analysis by professionals is not needed.

Description

technical field [0001] The invention relates to the technical field of identification of pathogenic bacteria, in particular to an Escherichia coli kit and an identification method. Background technique [0002] Escherichia coli is a Gram-negative bacillus, which belongs to the normal colony in the intestinal tract of animals. A small part of them can cause various local tissue infections such as the intestinal tract or urinary tract of humans or various animals under certain conditions, and is a common clinical infection. Pathogenic bacteria. The main clinical manifestations are severe watery diarrhea, bloody diarrhea, fever, abdominal cramps, and vomiting. In severe cases, acute kidney disease may occur. Children are at a higher risk of developing this complication. If not treated properly, it may be fatal. However, the drug resistance mechanism of Escherichia coli is complex and can be widely spread through plasmids, so it has natural resistance to some antibiotics, inclu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/10C12Q1/04C12R1/19
CPCC12Q1/10C12Q1/045G01N2333/245C12Q2334/52C12Q2334/70C12Q2334/10Y02A50/30
Inventor 蔡艳婷蔡克亚赵高岭刘美丽陈静曹洁茹李越峰牛亚静孙苛苛
Owner AUTOBIO DIAGNOSTICS CO LTD
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