SafePath Trichinae Immunoassay Kit 960-well
Product Code: TNP-960
Trichinellosis is the infection caused by the nematode Trichinella spiralis.
Although the nematode may be found in a wide variety of animals worldwide,
the domestic pig is the primary source of infection in humans in developed
Definitive diagnosis is by demonstration of the larvae in infected muscle tissue.
This approach is most successful when there is moderate to high worm burden
and maximum larval growth has occurred.
Recently, an excretory-secretory (ES) antigen has been purified from the
larvae of infected pigs. This antigen has a high degree of specificity for T.
spiralis. Antibody response will vary depending upon time of infection and
worm burden. A low worm burden will cause seroconversion between 28 to 42
days after initial infection. A high worm burden will cause seroconversion
between 14 to 28 days.
Principle of Procedure
During the first incubation, the antibodies in the sample binds to the ES antigens
in the test well. The next incubation then allows the anti-swine IgG peroxidase
complex to bind to the antigen-antibody complex. After a few washings to
remove unbound enzyme, a chromogen is added that develops a blue color in
the presence of the enzyme complex and peroxide. The stop solution ends the
reaction and turns the blue color to yellow.
Performance of Test
1 - Break off number of wells needed (two for controls plus number of
samples) and place in strip holder.
2 - Add 100 ul of negative control to well #1, 100 ul of positive control to well
#2, and 100 ul of the diluted test samples to the remaining wells.
Note: Negative and positive controls are supplied as prediluted. Use as is.
3 - Incubate at room temperature (15 to 25 ° C) for 10 minutes.
4 - Shake out contents and wash 3 times with diluted wash buffer.
5 - Add 2 drops (100 ul) of enzyme conjugate to each well.
6 - Incubate at room temperature for 10 minutes.
7 - Shake out contents and wash 3 times with wash buffer.
8 - Wash each well once with DI water.
9 - Add 1 drop (50 ul) of Substrate A and 1 drop (50 ul) of Substrate B to
every well. Mix by tapping strip sideways to vortex reagents. This will produce
complete mixing of the solutions.
10 - Incubate at room temperature for 10 minutes.
11 - Add 2 drops (100 ul) of stop solution.
12 - Zero ELISA reader on air, read wells at 450/620 to 650 nm or read
Each Kit contains 960 well