The document summarizes an ELISA kit for detecting Fasciola hepatica, a parasitic flatworm, in bovine serum or milk. The kit uses monoclonal antibodies coated on a microplate to detect antibodies to F. hepatica in samples. Testing takes 3 hours and provides reliable, specific results. A study using the kit found excellent concordance between serum and milk samples from the same animals.
The document summarizes an ELISA kit for detecting Fasciola hepatica, a parasitic flatworm, in bovine serum or milk. The kit uses monoclonal antibodies coated on a microplate to detect antibodies to F. hepatica in samples. Testing takes 3 hours and provides reliable, specific results. A study using the kit found excellent concordance between serum and milk samples from the same animals.
The document summarizes an ELISA kit for detecting Fasciola hepatica, a parasitic flatworm, in bovine serum or milk. The kit uses monoclonal antibodies coated on a microplate to detect antibodies to F. hepatica in samples. Testing takes 3 hours and provides reliable, specific results. A study using the kit found excellent concordance between serum and milk samples from the same animals.
The document summarizes an ELISA kit for detecting Fasciola hepatica, a parasitic flatworm, in bovine serum or milk. The kit uses monoclonal antibodies coated on a microplate to detect antibodies to F. hepatica in samples. Testing takes 3 hours and provides reliable, specific results. A study using the kit found excellent concordance between serum and milk samples from the same animals.
com FASCIOLA HEPATICA ELISA KIT For serum or milk (Bovine, Ovine) - Double wells - BIO K 211/2 - BIO K 211/5 Bovine fasciolosis caused by the digenic trematode Fas- Use of the kit ciola hepatica is a worldwide parasitic disease common in The kit is designed to follow ruminants. This two-host life cycle parasite is classically serological status of sera or found in farms where all conditions for the survival and milks the multiplication of the snail intermediate host ( Lym- nea truncatula) are fulfilled. This snail is mainly found in Reliable Results damp meadows (watering-places, brooks, springs,...). The use of monoclonal anti- Fasciola egg shedding occurs with faeces. Hat- body as conjugate ensures ching follows in water and gives rise to the miraci- excellent specificity and very dium which infests the snail. After multiplication reliable results. in this host, cercariae are eliminated and give rise The use of monoclonal anti- to infectious metacercariae fixed on a plant holder. bodies to purify one Fasciola Once ingested by a ruminant, young flukes mi- hepatica protein on the plate grate through the liver to reach bile ducts. The also makes it possible to ob- prepatent period is 8 to 10 weeks. Adults ap- tain an excellent specificity pear in the bile ducts and start to lay eggs. Liver damage and acute disease (especially in sheep) are caused by migrating immature parasites. Chro- nic disease occurs in cattle during the biliary phase. Ease-of-Use Minimal hands-on-time Zootechnical characteristics are hampered by the di- Room temperature incubation sease. Decrease in milk yield (-10%), weight loss, in- Results available in 140 termittent diarrhoea, anemia and fertility problems. minutes for single or batch Diagnosis of Fasciola hepatica in cattle can only be testing made after 8 to 10 weeks by coprological examination of faecal material. However, sometimes even repeated fecal examination cannot indentify any Fasciola hepati- ca infection due to the lack of sensitivity of this method. Acute distomatosis of the sheep is characterized by anemia and sometimes sudden mortality and chronic distomatosis by anemia, reduction of the dairy produc- tion, reduction of the average daily profit and oedemas
EIA Procedure
1- Microplate coated with monoclonal antibody and
Fasciola hepatica purified protein. 2- Add samples and positive control. Incubate 1 hour at 21°C+/-3°C. Wash 3- Add conjugate. Incubate 1 hour at 21°C+/-3°C. Wash 4- Add chromogen (TMB). Wait 10 minutes Add stop solution. Read at 450 nm
Bio-X Diagnostics - 38, Rue de la Calestienne (PAE) - 5580 Rochefort - Belgique
Tél : 0032(0)84.32.23.77 - Fax : 0032(0)84.31.52.63 - E-mail : a.ginter@biox.com www.biox.com Example of results 270 serum and 270 milk samples taken from the same animals were tested using the BIO K 211 kit. These samples came from twenty-seven Belgian farms. Their optical density readings were divided by the optimal density reading for the kit’s reference serum (S/P). Frequency histograms were then plotted for the blood sera (Graph 1) and milk samples (Graph 2).
Graph 1
sera
140
120
100
80 frequency
60
40
20 S/P E/P
0 0 0,4 0,8 1,2 -20 classes
Graph 2
Milks
120
100
80 frequency
60
40
20 S/P E/P
0 0 0,2 0,4 0,6 0,8 1 1,2 1,4 classes
Bio-X Diagnostics - 38, Rue de la Calestienne (PAE) - 5580 Rochefort - Belgique
Tél : 0032(0)84.32.23.77 - Fax : 0032(0)84.31.52.63 - E-mail : a.ginter@biox.com www.biox.com Example of results The frequency histograms generated for the 270 milk and 270 blood serum samples reveal that the thres- holds must be set preferentially at 20% of the kit’s reference serum’s signal. Graph 3 shows the correla- tion between the serological findings yielded by the blood sera and those yielded by the milk samples.
The concordance between the two tests is considered excellent. Landis et Koch, The measurement of observer agreement for categorical data Biometrics 1977, 33, 159-74
Bio-X Diagnostics - 38, Rue de la Calestienne (PAE) - 5580 Rochefort - Belgique
Tél : 0032(0)84.32.23.77 - Fax : 0032(0)84.31.52.63 - E-mail : a.ginter@biox.com www.biox.com Example of results
3 OD values obtanined with sera from sheep (6-month old) infected
experimentally with a single-dose of 5, 10, 20 or 40 metacercariae Delta from Fasciola hepatica .
2,5
2 10 Sheep
1,5
Infested Control
0,5
4 Sheep 0 0 2 4 6 8 10 12 14 16 18 20
Weeks Post
-0,5
Composition of the kit
BIO K 211 Fasciola Hepatica ELISA kit
BIO K 211/2 BIO K 211/5
Microplate 2 (96 tests) 5 (240 tests) Washing solution 1 X 100 ml (20 X) 1 X 250 ml (20 X) Dilution buffer 1 X 50 ml (5 X) 1 X 100 ml (5 X) Conjugate 1 X 0.5 ml (1 X) 1 X 1.4 ml (1 X) Positive serum 1 X 0.5 ml (1 X) 1 X 0.5 ml (1 X) Single component TMB 1 X 25 ml (1 X) 1 X 55 ml (1 X) Stopping solution 1 X 15 ml (1 X) 1 X 30 ml (1 X)
Bio-X Diagnostics - 38, Rue de la Calestienne (PAE) - 5580 Rochefort - Belgique
