Method for detecting neomycin B content in milk and application thereof

A technology of neomycin B and neomycin, applied in biochemical equipment and methods, biological testing, material inspection products, etc., can solve the problems of slow analysis speed, high detection cost, expensive detection instruments, etc., and achieve easy operation , highly specific effect

Inactive Publication Date: 2016-07-06
SHENZHEN ELDERLY MEDICAL RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the above method has high accuracy, it requires expensive detection instruments, high detection cost, and slow analysis speed, so it is mainly used in detection laboratories.

Method used

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  • Method for detecting neomycin B content in milk and application thereof
  • Method for detecting neomycin B content in milk and application thereof
  • Method for detecting neomycin B content in milk and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, the preparation of the nucleic acid aptamer that detects neomycin B (NeomycinB) in milk

[0041] 1. Preparation of nucleic acid aptamers

[0042] The RNA aptamer (rGrGrArCrUrGrGrGrGrCrGrArGrArArGrUrUrUrArGrGrUrCrC, wherein rG represents the G on the RNA) in the sequence capable of binding to neomycin B is truncated into two parts in its segment rA and rG, and one part is in its sequence The 3' end is modified with 15 thymine (T) spacers and 12 adenine (A) tails, named NEO1, and its nucleotide sequence is as follows: 5'-rGrArArGrUrUrUrArGrUrCrC-(T) 15 -(A) 12 -3' (sequence 1); another part modifies chemical fluorescent group (FAM) at its sequence 5' end, named after (NEO2), and its nucleotide sequence is 5'-FAM-rGrGrArCrUrGrGrGrGrCrGrA-3' (sequence 2 ).

[0043] Artificially synthesized NEO1 and NEO2 are the nucleic acid aptamers of neomycin B.

[0044] 2. Preparation of a kit for detecting neomycin B (NeomycinB) in milk

[0045] Preparation of nano-go...

Embodiment 2

[0053] Embodiment 2, detection of neomycin B (NeomycinB) in milk

[0054] 1. Sensitivity detection

[0055] The milk to be tested contains 250mMNaCl and 5mMMgCl 2 The 50mM Tris-HCl buffer solution was diluted 1:5 to obtain the test sample; stored at 4°C.

[0056] Take 284 μL of 10 nM nano-gold colloid solution linked to NEO1, 6 μL of 100 μM NEO2 solution, and 10 μL of detection sample prepared in Example 1 above, mix them and let stand at room temperature for 10 minutes to obtain a reaction solution.

[0057] Detect the fluorescence value of the reaction solution with a fluorescence detector; calculate the neomycin B content by the fluorescence value of the reaction solution;

[0058] The calculation formula is as follows: fluorescence value=-1.331×concentration+56.760.

[0059] The result is as figure 2 As shown, it can be seen that the concentration of neomycin B was detected linearly within 0.1 μM to 10 μM, and the lowest detection limit was 0.001 μM.

[0060] 2. Spec...

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Abstract

The invention discloses a method for detecting the content of neomycin B (Neomycin B) in milk and an application thereof. The present invention provides a complete set of nucleic acid aptamers for detecting neomycin B, consisting of nucleic acid aptamer NEO1 and nucleic acid aptamer NEO2; the nucleotide sequence of the nucleic acid aptamer NEO1 is sequence 1; the The nucleotide sequence of the nucleic acid aptamer NEO2 is sequence 2. Experiments of the present invention have proved that the invention overcomes the deficiencies in the prior art and provides a method for detecting neomycin B (Neomycin B) content in milk; Control and raw milk stations are used for on-site screening in large quantities.

Description

technical field [0001] The invention belongs to the field of food safety detection, in particular to a method for detecting the content of neomycin B (NeomycinB) in milk and an application thereof. Background technique [0002] Neomycin B is an aminoglycoside antibiotic, which is a quiescent bactericidal drug. Its mechanism of action is to pass through the cell membrane through active transport, irreversibly bind to one or several proteins of the bacterial ribosome 30S subunit, interfere with the formation of the initial complex between mRNA and the 30S subunit, and result in abnormal protein synthesis; abnormal protein binding into Bacterial cell membrane, resulting in cell membrane penetration and bacterial death. Neomycin is effective against aerobic Gram-negative bacilli and some Gram-positive bacteria. In Gram-negative bacilli, it has strong antibacterial activity against Escherichia coli and other Enterobacter species, Serratia spp., Proteus, and Morgan bacillus; et...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115G01N33/53
CPCC12N15/115C12N2310/16C12N2310/3517G01N33/53
Inventor 凌凯吴正治李富荣
Owner SHENZHEN ELDERLY MEDICAL RES INST
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