Vibrio parahaemolyticus immune colloidal gold rapid detection test strip
A hemolytic vibrio and test paper detection technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of detection precision and accuracy, low mortality rate, short validity period of test strips, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0046] Example 1 Preparation of anti-Vibrio parahaemolyticus monoclonal antibodies 3G9E9G3H7 and 3G9F7D5C9
[0047] 1. Preparation of immunogen and positive standard
[0048] Vibrio parahaemolyticus (ATCC No. 17802) was inoculated in 3% sodium chloride alkaline peptone water (APW), cultured with shaking at 37°C and 150r / min for 17 hours, counted, and inactivated by adding 0.3% formaldehyde solution at room temperature for 1 day. Adjust the concentration of Vibrio parahaemolyticus (ATCC No.17802) to 5×10 with normal saline 9 CFU / ml was used as immunogen; the concentration was adjusted to 10 with normal saline 8 cfu / ml was used as a positive control standard, and 3% sodium chloride alkaline peptone water (APW) was used as a negative control standard.
[0049] 2. Preparation of monoclonal antibodies
[0050] 1) Experimental animals: Three 8-week-old, weighing about 20 g female Balb / c mice were selected as experimental animals.
[0051] 2) Immunization method: each mouse was i...
Embodiment 2
[0076] Example 2 Characterization of Monoclonal Antibodies 3G9E9G3H7 and 3G9F7D5C9
[0077] 1. Monoclonal Antibody Subclass Identification
[0078] 1. Antigen coating: Coat goat anti-mouse secondary antibody IgG+A+M with 0.01MPBS, 50 μl per well, coat overnight at 4°C, discard the liquid in the well the next day, and wash the plate 3 times.
[0079] 2. Blocking: add 200 μl of 1% BSA to each well, and block overnight at 4°C. Pat the board dry the next day without washing it.
[0080] 3. Add monoclonal antibody hybridoma cell supernatant, 8 microwells for each sample, 50 μl per well. Incubate for 1 hour at 37°C.
[0081] 4. After washing the plate 4 times, add specific binding rabbit anti-mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, κ, λ, and incubate at 37°C for 1 hour.
[0082] 5. After washing the plate 4 times, add diluted horseradish peroxidase-labeled anti-rabbit secondary antibody IgG (H+L) to each well, and incubate at 37°C for 30 minutes.
[0083] After washing the pl...
Embodiment 3
[0096] Example 3 Reaction characteristic identification of monoclonal antibody 3G9E9G3H7 and 3G9F7D5C9 and Vibrio parahaemolyticus flagellin
[0097] Certainly
[0098] 1. Vibrio parahaemolyticus flagellin extraction:
[0099] (1) Vibrio parahemolyticus (ATCC17802) was inoculated in 3% sodium chloride alkaline peptone water (APW), cultured with shaking at 37°C and 150r / min for 17h, counted, and inactivated by adding 0.3% formaldehyde solution at room temperature for 24h .
[0100] (2) The next day, the collected bacteria were centrifuged and resuspended in pre-cooled 0.15M NaCl solution, placed in an ice bath for 15-30 minutes, homogenized at 1000 rpm for 45 seconds, and immediately placed on ice for 10 minutes.
[0101] (3) Centrifuge the homogenate at 10,000 rpm for 10 min at 4°C; take the supernatant and centrifuge at 16,000 rpm for 2 h at 4°C, discard the supernatant, and resuspend the pellet in TET (10 mM Tris, 2 mM EDTA, pH 8.0, 1% TritonX-100) buffer in the liquid. ...
PUM
| Property | Measurement | Unit |
|---|---|---|
| particle size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More