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目的对3株C型肉毒梭菌进行鉴定,并选择其中1株进行基因全序列测定。方法对3株C型肉毒梭菌(C6514、C314/91和CAXA)进行生化鉴定、检出试验、确证试验、毒力测定、毒素定型试验,并对C6514株进行神经毒素基因全序列测序。结果 3株菌均能发酵葡萄糖,不能发酵乳糖,不能发酵牛奶和形成吲哚;3株菌均为产毒株;C6514株毒力较高,为2×104 LD50/ml;加C型或混合型肉毒诊断血清或加明胶磷酸盐缓冲液(GPB)煮沸均可中和或灭活C型肉毒梭菌产生的毒素;BoNT/C测得的序列拼接后与GenBank中序列号为D90210的基因核苷酸和氨基酸序列同源性均为99%。结论 3株肉毒梭菌均为典型的C型肉毒梭菌。
Objective To identify three strains of Clostridium botulinum type C and choose one of them for the whole gene sequencing. Methods Three strains of Clostridium botulinum type C (C6514, C314 / 91 and CAXA) were biochemically identified, tested, confirmed, tested for virulence and toxin type. The complete sequence of the neurotoxin gene was sequenced in C6514 strain. Results All of the three strains could ferment glucose, lactose could not be fermented, milk could not be fermented and indole was formed. All three strains were producing strains. C6514 strain had a higher virulence of 2 × 104 LD50 / ml. Type botulinum diagnostic serum or boiled with gelatin phosphate buffer (GPB) can neutralize or inactivate the toxin produced by Clostridium botulinum type C. The sequence of BoNT / C is spliced and ligated with GenBank serial number D90210 Gene nucleotide and amino acid sequence homology was 99%. Conclusion All three strains of Clostridium botulinum are typical C. botulinum type C.