| 摘要: |
| 用包含16S-23S rDNA间隔区和23S rDNA的部分序列作为气肿疽特异性标志，以α毒素部分序列作为腐败梭菌特异性标志，建立了鉴别气肿疽梭菌和腐败梭菌疫苗生产用菌株的二重PCR方法。结果：气肿疽梭菌C54-1株扩增出大小为509bp的条带，腐败梭菌C55-1株、C55-16株均扩增出大小为148bp的条带，均与预期吻合；而产气荚膜梭菌C57-1株、C59-37株，肉毒梭菌C62-4株、诺维梭菌C61-4株均未扩增出任何条带。扩增产物的测序结果进一步证实了本方法的特异性。菌株的生物学特性试验结果也符合相应气肿疽梭菌和腐败梭菌的特点。结果表明，本研究所建立的二重PCR方法可用于气肿疽株和腐败梭菌疫苗生产用菌株的快速鉴定。 |
| 关键词: 气肿疽梭菌 腐败梭菌 16S-23S rDNA 二重PCR |
| DOI： |
| 投稿时间：2010-08-18修订日期：2010-08-18 |
| 基金项目: |
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| A Dual-PCR Assay for Differentiating theVaccine-producing Strains of C.chauvoei and C.septicum |
| pengxiaobing |
| (China Institue of Veterinary Drug Control) |
| Abstract: |
| A dual-PCR assay was developed to differentiate the vaccine-producing strains of C.chauvoei and C.septicum based on partial sequences containing the 16S-23S rDNA spacer region and 23S rDNA of C.chauvoei and partial alpha toxin gene of C.septicum. Strains of C.chauvoei C54-1, and C.septicum C55-1, C55-16 could be differentiated by this assay successfully. Size of 509bp was amplified from the strain C54-1, and 148bp from C55-1, C55-16. No specific band was amplified from control bacteria such as C.perfringens, C.botulinum, and C.novyi. Futhermore, specificity of PCR was confirmed by sequecncing of the amplified products and biological test The results indicated that dual-PCR could be a rapid way to differentiate the vaccine-producing strains of C.chauvoei and C.septicum. |
| Key words: C.chauvoei C.septicum 16S-23S rDNA dual-PCR |
