Detection card for rapidly and quantitatively detecting penicillin and application thereof
A quantitative detection and penicillin technology, applied in the field of penicillin detection, can solve problems such as low detection sensitivity of carlin, and achieve the effects of improving sensitivity, reducing production cost and shortening detection time.
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Embodiment 1
[0036] This embodiment is a preparation method of penicillin whole antigen, which preparation method comprises the following steps:
[0037] Weigh 5mg of penicillin G, add 1000μL of distilled water to dissolve, then add 30mg of EDC, mix well; place in a shaker at 25°C, shake at 180rpm for 1h to activate the carboxyl group;
[0038] Weigh 50mg of bovine serum albumin, add 2ml of 0.1M boric acid buffer solution with pH9.0;
[0039] The bovine serum albumin solution was added dropwise to the activated penicillin solution, placed in a shaker at 25° C., and shaken at 180 rpm for 2 hours to obtain the penicillin whole antigen.
[0040] The above-mentioned penicillin whole antigen was identified by ultraviolet scanning; the identification results were as follows: figure 1 shown;
[0041] Depend on figure 1 It can be seen that penicillin G has an absorption peak at 300nm, BSA and other proteins have absorption peaks at 280nm, and penicillin whole antigen has strong absorption peaks...
Embodiment 2
[0043] This embodiment is a method for labeling penicillin-binding protein with quantum dots, the method comprising the following steps:
[0044] Mix 1ml (1000μg) ZnS / CdSe composite quantum dots (produced by Shandong Ludu Biotechnology Co., Ltd., batch number LD028-L) and 1000μg N-hydroxysuccinimide, stir and react at room temperature for 20min, then add 2000μg N, N '-Dicyclohexylcarbodiimide and 2000 μg of penicillin-binding protein (from Shandong Lvdu Biotechnology Co., Ltd., product number LD-Ag089) were reacted at room temperature for 1.5 h, and the supernatant was discarded by centrifugation. Serum protein, 0.05% NaN 3 Dissolve in 0.01M PBS to obtain the quantum dot-labeled penicillin-binding protein.
Embodiment 3
[0046] This embodiment is a method for preparing a binding pad containing a quantum dot-labeled penicillin-binding protein, the preparation method comprising the following steps:
[0047] Select polyester fiber 6613 as the binding pad, soak the binding pad in pH7.4 0.01M PBS buffer containing 0.2% Triton X-100, 0.5% alanine, 0.05% NaN3 for 5min, take it out and dry it at 37°C; The penicillin-binding protein marked with quantum dots of 1 mg / ml (calculated by quantum dot mass) obtained in Example 2 was sprayed on the treated junction with a film-drawing instrument at a concentration of 10 μL / cm, and dried naturally at room temperature to form Conjugate pad containing quantum dot-labeled penicillin-binding protein.
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