Aptamer based methicillin-resistant staphylococcus aureus fluorescence detection method
A technology for methicillin-resistant Staphylococcus aureus, applied in the field of fluorescence detection of methicillin-resistant Staphylococcus aureus based on aptamers, can solve the problems of cumbersome preparation process, high price and high cost of monoclonal antibodies, and achieve easy preparation and labeling, increased sensitivity, and reduced cost
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Embodiment 1
[0042] (1) The magnetic nanoparticles used in the examples of the present invention are carboxyl-modified magnetic nanoparticles with a particle size of 800 nm, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd. PBP2a aptamer, synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., address: No. 698, Xiangmin Road, Songjiang District, Shanghai, preservation number: M23
[0043] The sequence of the PBP2a aptamer is:
[0044] 5'-CCATCCACACTCCGCAAGGGTGCCCCGGGGGGCTGTTCAGCGTGGTGGTGGGATGCCGTTTTGGTCCTTAGTCTCCGTCGTCGGCTGCCTCTACAT-3', the 5' end is labeled with the fluorescent group FITC;
[0045] The complementary DNA strand sequences of the aptamers are: 5'-GCGGAGTGTGGATGG-DABCYL-3'(Q-DNA-1),
[0046] 5'-CCTTGCGGAGTGTGG-DABCYL-3'(Q-DNA-2) and
[0047] 5'-CACCCTTGCGGAGTG-DABCYL-3'(Q-DNA-3), the 3' end is labeled with the quenching group DABCYL.
[0048] (2) In this example, based on the above-mentioned synthesized fluorescently labeled PBP2a aptamer and que...
Embodiment 2
[0068] (1) The magnetic nanoparticles used in the examples of the present invention are carboxyl-modified magnetic nanoparticles with a particle size of 800 nm, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd. PBP2a aptamer, synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., address: No. 698, Xiangmin Road, Songjiang District, Shanghai, preservation number: M25.
[0069] The sequence of the PBP2a aptamer is:
[0070] 5'-CCATCCACACTCCGCAAGGGTCGGTCTTCCCCTTCAGCTTGATGGGGTCCTGGGTGCCGGGATTTGGTTGCTGGTCTTCGTCGGCTGCCTCTACAT-3', the 5' end is labeled with the fluorescent group FITC; the complementary DNA strand sequence of the aptamer is: 5-CCTTGCGGAGTGTGG-DABCYL-3'(Q-DNA-2), the 3' end is labeled with a quenching group DABCYL.
[0071] (2) In this example, based on the above-mentioned synthesized fluorescently labeled PBP2a aptamer and quencher labeled aptamer complementary DNA strand, a highly sensitive and highly selective fluorescent detection syste...
Embodiment 3
[0084] (1) The magnetic nanoparticles used in the examples of the present invention are carboxyl-modified magnetic nanoparticles with a particle size of 800 nm, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd. PBP2a aptamer, synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., address: No. 698, Xiangmin Road, Songjiang District, Shanghai, preservation number: M23
[0085] The sequence of the PBP2a aptamer is:
[0086] 5'-CCATCCACACTCCGCAAGGGTGCCCCGGGGGGCTGTTCAGCGTGGTGGTGGGATGCCGTTTTGGTCCTTAGTCTCCGTCGTCGGCTGCCTCTACAT-3', the 5' end is labeled with the fluorescent group FITC;
[0087] The complementary DNA strand sequences of the aptamers are: 5'-GCGGAGTGTGGATGG-DABCYL-3'(Q-DNA-1),
[0088] 5'-CCTTGCGGAGTGTGG-DABCYL-3'(Q-DNA-2) and
[0089] 5'-CACCCTTGCGGAGTG-DABCYL-3'(Q-DNA-3), the 3' end is labeled with the quenching group DABCYL.
[0090] (2) In this example, based on the above-mentioned synthesized fluorescently labeled PBP2a aptamer and que...
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