Direct fluorescene immunoassay for viral antigens
A virus antigen and antigen technology, applied in the field of biological samples, can solve problems such as loss
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Embodiment I
[0256] [Example 1: LDFA detection of influenza A virus]
[0257] will repeat R- sv / cs cell culture monolayers with a series of 4 10-fold serial dilutions of influenza A virus (A / H3N2) or herpes simplex virus (HSV-1) (i.e., designated as samples: 4+, 3+, 2 + and 1+) inoculations and comparison with negative control (NC). Infected cultures were grown on coverslips inside shell flasks to allow virus replication for approximately 22 hours.
[0258] For each virus group, media was aspirated from 4+, 1+ and NC shell bottles. Phosphate buffered saline (PBS; 200 μl) was then added to each shell flask; and the cell monolayers were scraped coverslips and transferred to labeled 1.5 ml Eppendorf centrifuge tubes. Acetone (100%; 800 μl) was added to each centrifuge tube to a final volume of 1 ml to create an 80% acetone / cell suspension solution, which was incubated at room temperature for about 10 minutes to permeabilize the cells.
[0259] Permeabilized cells were then harvested and ...
Embodiment II
[0261] [Example II: Duet MAb virus screening in clinical aspirates]
[0262] Nasal discharge samples were collected from patients. Aliquot each sample into Tube A / B; Tube R / M; or Tube P / Ad (A-Influenza A; B-Influenza B; R-Respiratory Virus; M-Pneumovirus; P-Parainfluenza Virus ; Ad-adenovirus).
[0263] 1. Add 70 μl of cell suspension to each tube
[0264] 2. Add 2 drops of the corresponding MAb
[0265] 3. Incubate at 35℃~37℃ for 5 minutes
[0266] 4. Wash with 1.5ml of PBS
[0267] 5. Centrifuge at 2000×g for 2 minutes
[0268] 6. Aspirate the supernatant with a transfer pipette
[0269] 7. Add 20 μl of resuspension buffer to each tube
[0270] 8. Loading Slides
Embodiment III
[0271] [Example III: MAb cross-reactivity: list of materials]
[0272] The following is a list of the materials with lot numbers used in the cross-reactivity described in the studies presented in Examples IV and V herein
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