【摘要】 目的 对分离自感染性腹泻患者粪便的霍乱弧菌分离株(HL-1)和患者前期食用相同来源蟹体的霍乱弧菌分离株(HL-2)进行鉴定和全基因组测序分析。方法 对HL-1与HL-2进行分离培养、血清学鉴定、飞行时间质谱鉴定、实时荧光定量PCR鉴定、生化及耐药性测试和全基因组测序。依据基因组序列进行多位点序列分型(multi-locus sequence typing, MLST)、代谢通路、毒力基因、耐药基因分析。结果 确认HL-1和HL-2为非O1/O139群霍乱弧菌(non-O1/non-O139 Vibrio cholerae,NOVC),两者基因组相似度为98.85%,均不具有ctxAB和tcp毒力基因，在血平板上培养均产生明显β溶血环。分型结果显示HL-1为ST-684型，HL-2为ST-173型，与浙江省7株分离株同型。药敏实验结果显示，选取的17种药物中，HL-1仅对粘菌素和链霉素具有抗性；HL-2对其中7种具有抗性。结论 HL-1与HL-2在基因位点和表型上存在差异，HL-1倾向于孤立种群，HL-2可能来源于江浙地区。核酸检测和飞行时间质谱在细菌性感染突发事件的快速处置中具有独特优势。更多还原

【Abstract】 Objective To identify and perform whole genome sequencing analysis on a Vibrio cholerae isolate(HL-1) from feces of a patient with infectious diarrhea and a Vibrio cholerae isolate(HL-2) from crab of the same source consumed by the patient before onset. Methods We performed isolation and cultivation, serological identification, time-of-flight mass spectrometry identification, real-time fluorescence quantitative PCR identification, biochemical and antibiotic susceptibility test and whole genome sequencing on HL-1 and HL-2. Based on genome sequences, multi-locus sequence typing(MLST) and analysis of metabolic pathways, virulence genes and drug resistance genes were conducted. Results HL-1 and HL-2 were confirmed as non-O1/non-O139 Vibrio cholerae(NOVC), with a genomic similarity of 98.85%. Neither of them carried ctxAB and Tcp virulence genes. Both HL-1 and HL-2 produced obvious β-hemolysis rings on blood agar plates. The MLST results revealed that HL-1 belonged to ST-684, and HL-2 belonged to ST-173, which was the same type as 7 isolates from Zhejiang Province. The antibiotic susceptibility test results displayed that among the 17 selected drugs, HL-1 was only resistant to colistin and streptomycin, but HL-2 was resistant to 7 of them. Conclusion HL-1 and HL-2 differ in genetic loci and phenotypes. HL-1 tends to be an isolated strain, while HL-2 may originate from Jiangsu and Zhejiang regions. Nucleic acid testing and time-of-flight mass spectrometry have unique advantages in the rapid disposal of bacterial infection emergencies.更多还原