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Chemiluminiscence detection kit of streptococcus mutans and application method of chemiluminiscence detection kit

A technology for chemiluminescent detection and Streptococcus mutans, which is applied in the field of microbial detection, can solve the problems of time-consuming and laborious, inability to achieve high-sensitivity detection of Streptococcus mutans, and low sensitivity, and achieve rapid detection results

Inactive Publication Date: 2016-12-07
SOUTHWEST UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the traditional method based on the bacterial culture model has good reliability and high sensitivity, it is time-consuming and labor-intensive. It usually takes several days to get the results, and requires skilled professionals and a large number of laboratory instruments.
Although the PCR method based on bacterial nucleic acid has high specificity and can realize multi-component detection, it needs to go through complicated processes such as extracting nucleic acid and synthesizing specific primers.
In addition, the methods used to detect Streptococcus mutans also include Southern blot hybridization technology based on DNA probes and thiazolium blue colorimetric method. Sensitive detection

Method used

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  • Chemiluminiscence detection kit of streptococcus mutans and application method of chemiluminiscence detection kit
  • Chemiluminiscence detection kit of streptococcus mutans and application method of chemiluminiscence detection kit
  • Chemiluminiscence detection kit of streptococcus mutans and application method of chemiluminiscence detection kit

Examples

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Embodiment 1

[0026] Example 1 Preparation of Chemiluminescent Detection Kit for Streptococcus mutans

[0027] The chemiluminescent detection kit of Streptococcus mutans of this embodiment comprises the following components: immunoglobulin G-coupled magnetic beads, enzyme-labeled vancomycin, enzymatic chemiluminescent substrate, Streptococcus mutans suspension standard, buffers and washes.

[0028] The immunoglobulin G coupled magnetic beads are rat immunoglobulin G2a coupled magnetic beads, and its preparation method is: take 1.0 mL of carboxylated magnetic beads of 1.0 mg / mL, and use 2-( N -morpholine) ethanesulfonic acid (MES) buffer (0.01 mol / L, pH 5.5) was washed 3 times, and MES buffer (0.01 mol / L, pH 5.5) containing 40 mg EDC HCl and 10 mg NHS was added 1.0 mL, activation reaction for 15 minutes; the activated magnetic beads were washed 3 times with PBS buffer (0.01 mol / L, pH 7.4), and 2.0 mg / mL rat immunoglobulin G2a (solvent: 0.01 mol / L, pH 7.4) was added 7.4 PBS buffer) 1.0 mL, ...

Embodiment 2

[0034] Example 2 Use of the Chemiluminescent Detection Kit for Streptococcus mutans

[0035] Such as figure 1 As shown, the method for using the chemiluminescent detection kit of Streptococcus mutans prepared in Example 1 comprises the following steps:

[0036] (a) Dilute the Streptococcus mutans suspension standard with buffer to make a Streptococcus mutans working bacterial solution; add 10 µL of 2.0 mg / mL rat immunoglobulin G2a conjugate to 2.0 mL of Streptococcus mutans working bacterial solution Magnetic bead suspension (using buffer as solvent), incubate at 37°C for 1 hour, separate with magnet to obtain Streptococcus mutans-rat immunoglobulin G2a coupled magnetic bead complex, wash 3 times with washing solution, then add 500 µL 4.0 µg / mL alkaline phosphatase-labeled vancomycin solution (using buffer as solvent), incubate at 37°C for 1 hour, and separate with a magnet to obtain alkaline phosphatase-labeled vancomycin-streptococcus mutans-rat The immunoglobulin G2a-coup...

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Abstract

The invention discloses a chemiluminiscence detection kit of streptococcus mutans and an application method of the chemiluminiscence detection kit. The kit comprises immune globulin G coupling magnetic beads, enzyme-labeled vancomycin, an enzymatic chemiluminiscence substrate, a standard streptococcus mutans suspension, a buffer solution and washing liquid. The application method is characterized in that the vancomycin which is simple, cheap and stable is used as the molecular-recognition reagent, the vancomycin is used along with the specific molecular-recognition reagent immune globulin G to form a double-site molecular recognition mode, and the double-site molecular recognition mode is combined with the enrichment and separation technology and the enzymatic chemiluminiscence technology to perform whole-cell detection of the streptococcus mutans. The detection kit is fast in detection, convenient, accurate, specific, sensitive, stable and cheap, is hopefully applicable to the field detection and fast screening of the streptococcus mutans, and is capable of providing a powerful technical support platform for the streptococcus mutans detection in fields such as clinical diagnosis, food safety and environment detection.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and relates to a detection kit for Streptococcus mutans and a method for using the kit. Background technique [0002] Streptococcus mutans (Streptococcus mutans) is a Gram-positive anaerobic pathogenic bacteria that grows in the oral cavity. It has the characteristics of secreting acidic substances to corrode tooth enamel. It is one of the main cariogenic bacteria in humans. In addition, Streptococcus mutans can also cause subacute bacterial endocarditis. Therefore, the establishment of a rapid, convenient and sensitive detection method for Streptococcus mutans is of great significance to human health. [0003] Among the existing detection methods for Streptococcus mutans, the most mature method is the traditional method based on bacterial culture mode, and the most widely used method is the PCR method based on bacterial nucleic acid. Although the traditional method based on the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/543G01N21/76
CPCG01N21/763G01N33/54333G01N33/56944G01N33/6854G01N2800/18
Inventor 付志锋杨诗嘉
Owner SOUTHWEST UNIVERSITY
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