摘要
目的 建立一种可一次性检测多种腹泻致病菌的快速诊断方法。方法 根据原核生物 16SrDNA独特的序列特性 ,以 16SrDNA为模板 ,用PCR结合RDB方法来检测腹泻病原菌。结果 肠道常见致泻病原菌均可扩增出同样大小的片段 ,而特定的病原菌仅与其相应的探针杂交 ,敏感性试验可检测出约 12 0 μg/ml的DNA。 结论 利用原核生物 16SrDNA序列特异性而建立起来的PCR -RDB方法具有特异性高、重复性好、灵敏度高等优点 。
Aim To development a rapid diagnostic method for pathogens of diarrhea. Methods Based on unique sequence characterististics of 16S rDNA,the rDNA was frist amplified by polymerase chain reaction, then the amplified productds was hybridized to specific probe bound to Nylon Membrans. Hybrid product was detected by Dig Nuleic Acid Detection System.Results 16S rDNA could be amplified from multiple diarrhea pathogens. Only specific microbe could be hybridized with corresponding probe. It could detect DNA as few as 120μg/ml.Conclusion The PCR-RDB based on unique sequence characteristic of 16S rDNA is a method of high specificity,good reproducibility and high sensitivity. It' s a practical method for diarrhea pathogens.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第1期33-35,共3页
Chinese Journal of Zoonoses
