Colloidal gold test strip for detecting human surfactant protein A, as well aspreparation method and application of colloidal gold test strip
A surface-active protein, colloidal gold test paper technology, applied in the field of medical testing, can solve the problems of reducing the mortality rate of neonatal respiratory distress syndrome, and achieve the effects of both sensitivity and specificity, simple operation and short duration
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Embodiment 1
[0051] Embodiment 1 detects the colloidal gold test strip of human surfactant protein A
[0052] The colloidal gold test strip for detecting human surfactant protein A is composed of water-absorbing pad 7, nitrocellulose membrane 3, immune colloidal gold pad 2, sample pad 1 and polyvinyl chloride bottom plate 6 in sequence; the nitrocellulose membrane is wrapped with Quality control line C line 5 and detection line T line 4, C line is the secondary antibody, T line is rabbit anti-human surfactant protein A polyclonal antibody II; the immune colloidal gold pad is coated with colloidal gold labeled mouse antibody Glass fibers of human surfactant protein A monoclonal antibody I.
Embodiment 2
[0053] Embodiment 2: the preparation method of the colloidal gold test strip that detects human surfactant protein A
[0054] Preparation of colloidal gold:
[0055] 1. Add 48.5ml of deionized water into a 100ml round bottom flask, stir and boil, and use a clean condenser to reflux for 10min.
[0056] 2. Take 0.5ml of HAuCl respectively 4 The aqueous solution (1wt%) was added to 1ml of trisodium citrate aqueous solution (0.35-1.485wt%), stirred evenly to form a 2.5ml mixed solution, and then quickly added to the boiling water in step 1. It can be observed that the color of the solution changes rapidly from colorless to blue-gray, and finally turns into bright red through purple transition.
[0057] 3. The reaction solution was heated, stirred and refluxed for 1 hour, then the heating was stopped, and the temperature was naturally lowered to room temperature while stirring.
[0058] Preparation of colloidal gold labeling complex:
[0059] 1. Add 1ml of colloidal gold soluti...
Embodiment 3
[0079] Embodiment 3: the sensitivity analysis of the colloidal gold test paper strip prepared by embodiment 2
[0080] Get 25 colloidal gold test strips for the detection of human surfactant protein A made in the above-mentioned embodiment 2. Dissolve the human surfactant protein A antigen gradient in the standard diluent, the concentrations are 1000ng / ml, 500ng / ml, 400ng / ml, 200ng / ml, 0ng / ml, test 5 strips for each sample concentration, and the test time is 10 minute. Table 1 shows the visual interpretation results. The results show that when the content of human surfactant protein A in the detection sample solution is lower than 200ng / ml, the test strip of the present invention does not develop color, that is, the detection rate is 0. When the content of human surfactant protein A in the detection sample solution is more than or equal to 400ng / ml, the test strip can be detected and interpreted normally, and the detection rate is 100%. The results show that the colloidal g...
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