Development of an immunodetection test for a botulinum-like neurotoxin produced by Clostridium sp. RKD.

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dc.contributor.authorDixit, Aparnaen_US
dc.contributor.authorAlam, S Ien_US
dc.contributor.authorDhaked, R Ken_US
dc.contributor.authorSingh, Lokendraen_US
dc.date.accessioned2006-09-07en_US
dc.date.accessioned2009-05-27T07:33:59Z
dc.date.available2006-09-07en_US
dc.date.available2009-05-27T07:33:59Z
dc.date.issued2006-09-07en_US
dc.description.abstractBACKGROUND & OBJECTIVES: Clostridial neurotoxins are among the most toxic substances known and cause severe illnesses in both humans and animals. A neurotoxigenic Clostridium sp. (strain RKD) isolated from intestine of decaying fish produced a novel, botulinum type B like neurotoxin as suggested by mouse bioassay, protection with anti-botulinum antibodies and PCR. The aim of the present investigation was to develop a laboratory based detection assay as an alternative to the mouse bioassay without compromising sensitivity and specificity. METHODS: Growth and toxin production were carried out in trypticase peptone yeast-extract glucose (TPYG) broth. Toxicity was estimated in terms of minimum lethal dose (MLD) by mouse bioassay. The toxin was partially purified by acid precipitation. It was used for toxoid preparation by formaldehyde treatment. This purified IgG was used for detection of neurotoxin using indirect ELISA. The culture supernatant was concentrated using a stirred cell with a 50 kDa cut-off membrane at 4 degrees C. Further purification was carried out using Prep cell. Fractions showing toxicity and sufficient purity were pooled, concentrated and analyzed on sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The toxin was purified with a recovery of 8.56 per cent. Polyclonal antiserum was raised in mice using partially purified toxin with a titre of 1: 80000. A detection assay with sensitivity of approximately 15 and 300 ng/ml for partially purified and crude toxins, respectively were achieved using an indirect ELISA method. INTERPRETATION & CONCLUSION: The Clostridium sp. RKD produced a potent neurotoxin earlier shown to have novelties. A specific detection assay for the neurotoxin has been developed that may be useful both from food safety and clinical point of view.en_US
dc.description.affiliationBiotechnology Division, Defence Research and Development Establishment, Gwalior, India.en_US
dc.identifier.citationDixit A, Alam SI, Dhaked RK, Singh L. Development of an immunodetection test for a botulinum-like neurotoxin produced by Clostridium sp. RKD. Indian Journal of Medical Research. 2006 Sep; 124(3): 355-62en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/21212
dc.language.isoengen_US
dc.source.urihttps://icmr.nic.in/ijmr/ijmr.htmen_US
dc.subject.meshAnimalsen_US
dc.subject.meshBiological Assayen_US
dc.subject.meshBotulinum Toxins --isolation & purificationen_US
dc.subject.meshClostridium --chemistryen_US
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assay --methodsen_US
dc.subject.meshFishes --microbiologyen_US
dc.subject.meshImmunoglobulin G --metabolismen_US
dc.subject.meshMiceen_US
dc.subject.meshNeurotoxins --isolation & purificationen_US
dc.titleDevelopment of an immunodetection test for a botulinum-like neurotoxin produced by Clostridium sp. RKD.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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