A method for detecting benzyl penicillin

A technology of benzyl penicillin and centrifuge tubes, applied in the detection field, can solve the problems of high cost, low specificity and sensitivity, and achieve the effects of fast reaction speed, improved sensitivity and low process cost

Active Publication Date: 2018-10-19
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problems of relatively low specificity and sensitivity and high cost of the method for detecting benzyl penicillin in the above prior art, the present invention provides a target-induced nucleic acid with high specificity and sensitivity, low cost and fast detection speed. Method for detection of benzyl penicillin by conformational change of aptamer

Method used

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  • A method for detecting benzyl penicillin
  • A method for detecting benzyl penicillin
  • A method for detecting benzyl penicillin

Examples

Experimental program
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preparation example Construction

[0041] The preparation method of described biosensor comprises the following steps:

[0042] (1) Pretreatment of DNA strands;

[0043] (2) Mixing in a homogeneous solution;

[0044] (3) The chromogenic agent ABTS is mixed with the mixed solution for detection.

[0045] Described preparation method, specific operation steps are as follows:

[0046] (1) Put the centrifuge tube containing the DNA chain into a centrifuge at 12000 R / min, and centrifuge for 10 minutes. After centrifugation, add an appropriate amount of sterilized water according to the ratio on the centrifuge tube, and then place it in a shaker for 30 seconds. , so that the chain fusion uniform.

[0047] (2) Add sterilized water, 10× buffer buffer, HAP1 and the target object to be tested into the centrifuge tube, shake for 30s, and incubate in a 37°C incubator for 1h;

[0048] (3) Take out the centrifuge tube incubated in step 2, add S1 and S2, phi29 DNA polymerase, and dNTPs into the centrifuge tube, shake for ...

Embodiment 1

[0053] The main steps of the specific operation process are as follows:

[0054] a. Put the centrifuge tube containing the DNA chain into a centrifuge at 12000 R / min, and centrifuge for 10 minutes. After the centrifugation is completed, add an appropriate amount of sterilized water according to the ratio on the centrifuge tube, and then place it in a shaker for 30 seconds. Blend the strands evenly.

[0055] b. Add sterilized water, 10× buffer, HAP1 (10 μM), and the target object to be tested (10 μM) into a centrifuge tube, shake for 30 seconds, and incubate in a 37°C incubator for 1 hour.

[0056] c. Take out the centrifuge tube incubated in b, and add different concentrations of S1 (2 μM, 5 μM, 10 μM, 15 μM) and S2 (5 μM), phi29 DNA polymerase (2 μL), dNTPs (2 μL) into this In a centrifuge tube, shake for 30s, and incubate in a 37°C incubator for 1h;

[0057] d. Take out the centrifuge tube incubated in (c), add HAP2 (5μM) and Nt.BbvCI (2μL) endonuclease into the centrifuge...

Embodiment 2

[0064] The main steps of the experimental process are as follows:

[0065] a. Put the centrifuge tube containing the DNA chain into a centrifuge at 12000 R / min, and centrifuge for 10 minutes. After the centrifugation is completed, add an appropriate amount of sterilized water according to the ratio on the centrifuge tube, and then place it in a shaker for 30 seconds. Blend the strands evenly.

[0066] b. Sterilized water, 10× buffer buffer, HAP1 (10 μM), target substances to be tested benzicillin (10 μM), streptomycin (10 μM), oxytetracycline (10 μM), Kanamycin Add protein (10 μM) into the centrifuge tube, shake for 30 seconds, and incubate for 1 hour in a 37°C incubator.

[0067] c. Take out the centrifuge tube incubated in b, then add different concentrations of S1 (10 μM) and S2 (5 μM), phi29 DNA polymerase (2 μL), dNTPs (2 μL) into the centrifuge tube, shake for 30 seconds , and incubated in an incubator at 37°C for 1 hour;

[0068] d. Take out the centrifuge tube incub...

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Abstract

The invention provides a method for detecting benzyl penicillin. The detection of the benzyl penicillin is realized in a homogeneous solution; amplification of a signal is realized in a two-step circulating manner, so that high-sensitivity detection of the benzyl penicillin is realized, and a relatively small lower detection limit is obtained.

Description

technical field [0001] The invention relates to a detection method, in particular to a method for detecting benzyl penicillin based on target-induced conformational changes of nucleic acid aptamers. Background technique [0002] Benzyl penicillin is a kind of antibiotic, which is extracted from Penicillium, because the molecule contains penicillin, it can destroy the cell wall of bacteria and play a bactericidal role in the reproduction period of bacterial cells. Penicillin is less toxic to the human body, but it can cause severe allergic reactions. The probability of allergic reactions is 5%-10%. Symptoms include dyspnea, cyanosis, drop in blood pressure, coma, rigid limbs, and finally convulsions. Failure to rescue in time can cause death . Moreover, the occurrence of allergic reactions has nothing to do with the size of the drug dose. For patients who are allergic to penicillin, even a small amount can cause shock. The currently reported detection methods for benzyl pen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
CPCG01N33/5308G01N2415/00
Inventor 王玉崔雪君黄加栋刘素裴倩倩冷雪琪张雪宋晓蕾王敬锋
Owner UNIV OF JINAN
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