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4 , reagent storage <br> stored at 2-8 ° C, the diluted washing solution can be stored at room temperature for one week name specification Microporous strip Coated Legionella antigens (1-6 groups) 12 X 8 wells Enzyme complex 1×15ml/bottle labeled HRP goat anti-human Legionella IgG/M/A antibody white cover Positive control 1×0.35ml/bottle red cover Standard 1 x 0.5 ml / bottle blue cover Negative control 1 x 0.35 ml / bottle green cover Sample diluent 1 x 30ml / bottle Substrate coloring solution 1×15ml/bottle containing TMB Concentrated washing solution (10X) 1 x 100ml / bottle Stop solution 1 × 15ml / bottle of 1M sulfuric acid
5 , sample collection and preparation <br> venipuncture to take blood, to avoid cross-infection, prohibit the use of hemolysis, contaminate blood samples. The blood sample should not be stored for more than 8 hours at room temperature, and should not be stored for more than 48 hours at 2-8 °C. If it needs to be stored for a longer period of time, it should be stored at -20 °C or lower.
6. Materials and equipment required for the experiment, but this kit does not provide <br> microplatette micropipette 10ul and 200ul
Multichannel pipette (50-200ul)
Deionized water plastic bottle washing sample gun measuring paper towel
7. Experimental procedure (1), equilibrate the components in the kit to room temperature.
(2) Determine the required microporous strip to fix it on the plate holder. (1 blank hole, 1 negative control hole, 3 standard holes, 1 positive control hole)
(3) Diluted according to 1:21 sample, negative control, standard, positive control (10ml serum + 200ml sample dilution)
(4) Add 100 ul of sample dilution solution to the A1 well as a blank well, and add 100 ul of diluted quality control, standards and samples to each well.
(6), room temperature (20-25 ° C) reaction 25 ± 5 minutes (7), wash the plate 5 times (8), add 100ul of enzyme conjugate (9) per well, room temperature (20-25 ° C) reaction 25 ± 5 minutes (10), wash plate 5 times (11), add 100 ul of substrate solution (12) per well, react at room temperature for 10-15 minutes (13), add 50 ul of stop solution (14) per well, within 30 minutes. Read the results of the microplate reader or read at 450nm/620-650nm
Note: wash repeatedly five times during washing
8 , the results of the description
CUTOFF OD value = reference product OD x correlation factor (CF)
The CF value is indicated on each kit.
Negative sample: sample OD/CUTOFF OD ≤ 0.90
Gray area: 0.91<sample OD/CUTOFF OD<1.09
Positive sample: sample OD/CUTOFF OD≥ 1.10
This translation is for reference only, please refer to the original for details.
Legionella antibody ELISA test kit instruction manual
Legionella antibody ELISA test kit
(US DRG original import: item number, EIA2814; specifications, 96T)
1. Scope of application <br> As an auxiliary means for qualitative detection of total anti-M. striata in human serum, it is an in vitro diagnostic reagent. It is only used for research in the United States.
2 , the experimental principle <br> microplate is coated with activated Legionella antigen, the experimental process consists of three steps
A. The reaction of the serum to be tested in the coated plate. Specific antibodies in the serum can be combined with the antigen in the coated plate, and the unbound sample is removed by washing.
B. Add enzyme-linked goat anti-human IgA/M/G, and the enzyme conjugate can react with the antibody bound to the coated plate in the previous step, and wash the plate.
C, the substrate liquid color reaction, through the color change, the content of the antibody can be judged.
3 , reagent composition