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〔目的〕探讨16S rDNA序列分析法在国境口岸监测外来病原微生物的可行性。〔方法〕按照0.5 mg/0.5 ml样品接种10 ml增菌液的比例将压载舱淤泥样品分别接种到营养肉汤和碱性蛋白胨水中,37℃过夜,震荡摇匀。增菌后划线接种在选择培养基上,过夜培养后挑取阳性菌落纯培养,按照试剂盒说明提取细菌基因组,扩增细菌基因组16S rDNA片段,将扩增产物纯化后进行序列分析。〔结果〕从48份样品中共检测12种细菌,其中检测到的霍乱弧菌有1株为B33株。这株霍乱弧菌最早在2004年分离自非洲东南部国家莫桑比克的贝拉港,是O1群霍乱弧菌的经典型与Eltor型杂交后产生的菌株,O139群霍乱弧菌尚未检测到。〔结论〕16S rDNA序列分析法可用作国境口岸外来微生物的监测和分析。
[Objective] To explore the feasibility of 16S rDNA sequence analysis in monitoring foreign pathogenic microorganisms in frontier ports. [Method] The ballast tank sludge samples were respectively inoculated into nutrient broth and alkaline peptone water according to the proportion of 0.5 ml / 0.5 ml sample inoculated with 10 ml of broth. After shaking at 37 ° C overnight, shake well. Strains were inoculated on the selection medium after enrichment and cultured overnight. The positive colonies were picked and purified. The bacterial genome was extracted according to the kit instructions. The bacterial genome 16S rDNA fragment was amplified. The amplified product was purified and sequenced. [Results] A total of 12 kinds of bacteria were detected from 48 samples, of which 1 strain of Vibrio cholerae was B33. This strain of V. cholerae was isolated from Beira port in Mozambique in southeastern Africa in 2004 as the first strain of V. cholerae O1 group that produced after the hybridization with Eltor type. Vibrio cholerae O139 strain has not been detected yet. [Conclusion] The 16S rDNA sequence analysis method can be used as the monitoring and analysis of the foreign microorganisms at the border crossings.