A fluorescent detection method for isocarbophos and profenofos based on nucleic acid aptamer

A nucleic acid aptamer and isocarbophos technology, which is applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems in the field of food safety in its infancy, and achieve high sensitivity and selectivity

Active Publication Date: 2017-06-23
ANHUI SCI & TECH UNIV
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Problems solved by technology

[0004] At present, the research on nucleic acid aptamer biomimetic molecular recognition system has received extensive attention both internationally and domestically, and relevant research progress has emerged continuously, but its application in the field of food safety is still in its infancy

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  • A fluorescent detection method for isocarbophos and profenofos based on nucleic acid aptamer
  • A fluorescent detection method for isocarbophos and profenofos based on nucleic acid aptamer
  • A fluorescent detection method for isocarbophos and profenofos based on nucleic acid aptamer

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Embodiment Construction

[0030] The present invention will be further described below in conjunction with accompanying drawing and embodiment, but the present invention is not limited to embodiment, under the premise of not departing from the gist of the present invention, any improvement done all falls within the protection scope of the present invention.

[0031] Fluorescence detection method of isocarbophos and profenofos based on nucleic acid aptamer, please refer to figure 1 ,Specific steps are as follows:

[0032] 1), hybridizing the nucleic acid aptamer F-ssDNA with the complementary sequence Q-B.

[0033] The nucleic acid aptamer F-ssDNA is a 35-base single-stranded DNA that recognizes isocarbophos and profenofos with the fluorescent group FAM labeled at the 5′ end, and the sequence is:

[0034] 5'-FAM-AGCTTGCTGCAGCGATTCTTGATCGCCACAGAGCT-3'.

[0035] The complementary sequence Q-B is a 15-base single-stranded DNA marked with DABCYL at the 3' end, and the sequence is:

[0036] 5'-GAATCGCTGCA...

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Abstract

The invention relates to the field of biotechnology, in particular to a nucleic acid aptamer-based fluorescence detection method for isocarbophos and profenofos. The detection principle is to label the 5′ end of the nucleic acid aptamer with a fluorescent group, design a sequence that hybridizes complementary to the nucleic acid aptamer and mark the quenching group at the 3′ end, and the nucleic acid aptamer will fluoresce when it hybridizes with the complementary sequence. The signal is very weak. When isocarbophos and profenofos recognize and bind to the nucleic acid aptamer, the complementary sequence will be dissociated and the fluorescence signal will be enhanced. The change of the fluorescence signal is used to establish a standard curve to determine the minimum detection limit and linear range. According to the standard The curve determines the content of the target molecule in the sample. The method is fast and simple, has high sensitivity and selectivity, and is suitable for the rapid detection of isocarbophos and profenofos in samples.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid aptamer-based fluorescence detection method for isocarbophos and profenofos. Background technique [0002] Organophosphorus pesticides are the main pesticides in my country, and the national standard stipulates that the maximum allowable residue of organophosphorus pesticides in agricultural products is 0.2mg / kg (GB / T5009.199-2003). Isocarbophos is a highly toxic and banned pesticide, and profenofos is an organophosphorus pesticide used in limited quantities. However, in recent years, due to the unreasonable use of pesticides, pesticide residues in agricultural products have seriously exceeded the standard, and food poisoning incidents have gradually increased, which has seriously endangered my country's food safety. The poisonous cowpea incident in 2010 was caused by the excessive use of the highly toxic pesticide isacarbophos. The monitoring of pesticide residues i...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 王丽徐龙峰王蓓蓓王丹丹吴丛婷
Owner ANHUI SCI & TECH UNIV
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