A型肉毒神经毒素基因的PCR检测

目的:建立快速筛查A型肉毒毒素的PCR方法。方法:根据GenBank中报道的肉毒毒素基因序列,综合应用多种生物软件分析设计特异的检测引物,从提取的基因组DNA、热裂解产物和菌液等不同形式的模板中扩增大小为457bp的A型肉毒毒素特异基因片段,以肉毒梭菌其他血清型及破伤风梭菌为对照。结果:检测方法无交叉反应,灵敏度可达10pgDNA,3×103个菌。结论:建立的检测方法特异性强、灵敏度高,可以用于A型肉毒毒素基因的快速筛查。

Detection of the Gene Encoding Botulinum Neurotoxin Type A by PCR

Objective The polymerase chain reaction PCR was used as the basis for the development of highly sensitive and specific diagnostic detection for organisms harboring botulinum neurotoxin type A gene. Methods Synthetic DNA primers were selected from nucleic acid sequence data for Clostridium botulinum neurotoxin type A. The PCR was adjusted for optimal amplification of the target fragment. The sensitivity of the detection was determined with different concentrations of genomic DNA and strains from strains producing different botulinum neurotoxin types. Results As little as 10 pg of DNA approximately clostridial strains was detected. Other botulinum neurotoxin types were also detected by the PCR and they all showed negative. Conclusion The PCR system is specific and sensitive for the identification of botulinum neurotoxin type A.