Clostridium Botulinum
Clostridium Botulinum is a single celled bacteria and has organelles, not bounded by a membrane. They are asexual and reproduce by binary fission. It belongs to Kingdom Bacteria because it has peptidoglycan in its cell wall. It is unicellular and lack nucleus. It is obligate anaerobe as well as have ability to produce by spore so it falls under Phylum Firmicutes. It is classified in class Clostridia and further in order Clostridiales due to its spore forming ability. It is in Genus Clostridium because of being gram positive rod shaped(bacillus). It is originally founded in blood sausage and Latin word for sausage is botulism so is under Species Clostridium. This bacterium requires pH …show more content…
Needs more sensitivity and rapidity
3. Test can work under range of only some sample forms such as faces, food and environmental samples.
4. Animal welfare issues
New advanced method for isolating and identifying Amplified ELISA test
Method-
Preparation of sample – inoculate food samples from agar plates into TPGY. And adjust ph between 7.4-7.6.
Preparation of microtiter plates- coat each well of microtiter plate with 100µl of dilution of goat type in bicarbonate buffer. Store plate with coating buffer overnight at 4°c.
ELISA analysis of culture media
a. Remove plate from 4°c storage and wash plate 5 times in tri buffered saline (TBST). Also block plate in casein buffer by filling all wells to top and incubate at 35°c for 90 minutes. Make negative and positive controls.
b. Wash the blocked plate as above and add toxic samples and controls
c. Incubated toxic containing samples store at 35°c for 2 hours. Add diluted biotin-labelled goat antibody and incubate for 60min at 35°c.
d. Wash it again with TSBT.
e. Add streptavidin- alkaline phosphate conjugate diluted 1:10,000 in casein buffer and incubate for 60min at 35°c.
f. Add 50µl of GIBCO substrate solution. And read absorbance at
Clostridium Botulinum is a single celled bacteria and has organelles, not bounded by a membrane. They are asexual and reproduce by binary fission. It belongs to Kingdom Bacteria because it has peptidoglycan in its cell wall. It is unicellular and lack nucleus. It is obligate anaerobe as well as have ability to produce by spore so it falls under Phylum Firmicutes. It is classified in class Clostridia and further in order Clostridiales due to its spore forming ability. It is in Genus Clostridium because of being gram positive rod shaped(bacillus). It is originally founded in blood sausage and Latin word for sausage is botulism so is under Species Clostridium. This bacterium requires pH …show more content…
Needs more sensitivity and rapidity
3. Test can work under range of only some sample forms such as faces, food and environmental samples.
4. Animal welfare issues
New advanced method for isolating and identifying Amplified ELISA test
Method-
Preparation of sample – inoculate food samples from agar plates into TPGY. And adjust ph between 7.4-7.6.
Preparation of microtiter plates- coat each well of microtiter plate with 100µl of dilution of goat type in bicarbonate buffer. Store plate with coating buffer overnight at 4°c.
ELISA analysis of culture media
a. Remove plate from 4°c storage and wash plate 5 times in tri buffered saline (TBST). Also block plate in casein buffer by filling all wells to top and incubate at 35°c for 90 minutes. Make negative and positive controls.
b. Wash the blocked plate as above and add toxic samples and controls
c. Incubated toxic containing samples store at 35°c for 2 hours. Add diluted biotin-labelled goat antibody and incubate for 60min at 35°c.
d. Wash it again with TSBT.
e. Add streptavidin- alkaline phosphate conjugate diluted 1:10,000 in casein buffer and incubate for 60min at 35°c.
f. Add 50µl of GIBCO substrate solution. And read absorbance at