Anisakidae ELISA Kit

Intended Use:
The Bordier Anisakidae ELISA kit is intended for the quantitative detection of IgG antibodies against parasites of Anisakidae family in human serum. Serology is an aid for diagnosis and cannot be used as the sole method of diagnosis.

Background:
Anisakidosis is a worldwide helminthic zoonosis caused by some sea fish nematodes of the family Anisakidae. Humans can be infected by ingesting raw or undercooked contaminated fish meat. Adult worms of anisakid nematodes reside in the digestive tract of marine mammals (final hosts). After being embryonated in the water, the eggs release infesting L3 larvae into the water, which are ingested by the planktonic crustacean (intermediate host). After consumption by fish or cephalopods (paratenic hosts), the larvae migrate into the viscera and the peritoneal cavity so that they no longer evolve. After ingestion by marine mammals, the larvae develop into adult worms. Man, accidental host, is a parasitic dead end. Most infected people do not show any symptoms. However, in some cases, symptoms appear at the gastric stage (epigastric pain, nausea and vomiting), intestinal (abdominal pain, nausea, vomiting and diarrhea) or at the allergic stage (hives, pruritus, angioedema and bronchospasm). Diagnosis is based on signs and symptoms plus a history of exposure and a positive result by serological testing.

Principle and Presentation:
The kit provides all the material needed to perform 96 enzyme-linked immunosorbent assays (ELISA) on breakable microtitration wells sensitized with Anisakidae excreted/secreted (E/S) larval antigens. Specific antibodies in the sample will bind to these antigens and washing will remove unspecific antibodies. The presence of parasite specific antibodies is detected with a Protein A – alkaline phosphatase conjugate. A second washing step will remove unbound conjugate. Revealing bound antibodies is made by the addition of pNPP substrate which turns yellow in the presence of alkaline phosphatase. Color intensity is proportional to the amount of Anisakidae specific antibodies in the sample. Potassium phosphate is added to stop the reaction. Absorbance at 405 nm is read using an ELISA microplate reader. The test can be performed with automatic systems, but this must be validated by the user.