多重PCR技术在调查分析水族馆海兽饲养环境中致病菌的应用

Application of Multiplex PCR Technique in Diagnosis of Pathogenic Bacteria in Marine Animal Breeding Environment of Aquarium

为了利用多重PCR技术快速准确地分析出水族馆海兽饲养区域内所存在的细菌种类与数量,为进一步加强疾病防控提供重要依据.本文对安徽省某水族馆海洋哺乳动物饲养区域进行前后两次采样和细菌增殖培养,通过提取核酸后使用通用引物对模板进行多重PCR检测,再经16S rRNA测序,经Gene Bank数据库比对,结果显示,两次分别检测出13种不同微生物,包括致病菌、腐败菌、环境菌等,分布在海兽馆各区域.表明多重PCR技术对该环境致病菌分离鉴定较为快速有效.

Multiplex PCR technology is used to quickly and accurately analyze the bacteria species and quantity in the marine animal breeding area of aquarium which will provide important basis for disease prevention and control.The collected cotton swab samples are cultured twice for bacterial proliferation,extracted nucleic acid and detected by multiplex PCR with universal primers.Finally,the results are sequenced by 16S rRNA and compared with gene bank database.13different microorganisms were detected twice,including pathogenic bacteria,spoilage bacteria,environmental bacteria distributed in the areas everywhere.Through twice sampling and detection of marine mammal breeding area in an aquarium in Anhui Province,it is found that this method is more rapid and effective in practical application.