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目的通过对泛耐药肺炎克雷伯菌JM45的质粒1(p JM45-1)耐药元件分析,从基因组水平研究其泛耐药的遗传学基础。方法采用第二代高通量测序技术平台进行全基因组测序,生物信息学方法分析p JM45-1质粒的耐药元件,并与已在NCBI登录的质粒序列作聚类分析(Fast Minimum Evolutin法)。结果 p JM45-1质粒大小为317 156 bp,功能注释分析发现该质粒为可接合转移质粒,携带了抗菌药物耐药编码基因、重金属离子耐受编码基因、毒素编码基因和转座子以及插入序列等83个耐药相关编码基因。p JM45-1质粒与耐药质粒R100(登录号:AP000342.1)的全长94 281个序列中的45 995个序列有99%相同;与接合性质粒F质粒(登录号:AP001918.1)的全长99 159个序列中的8322个序列有87%相同。p JM45-1质粒与源自肺炎克雷伯菌的质粒在同一簇(cluster),与源自其他肠杆菌的质粒不在一个簇。结论肺炎克雷伯菌JM45 p JM45-1质粒携带大量耐药元件,是该菌株进化为泛耐药的主要原因。p JM45-1是可接合转移质粒,可将耐药基因在细菌间进行水平转移,造成耐药菌的播散。
OBJECTIVE: To study the genetic basis of pan-drug resistance at the genome level by analyzing the resistance element of plasmid 1 (p JM45-1) of pan-resistant Klebsiella pneumoniae JM45. Methods Genome-wide sequencing was carried out using the second-generation high-throughput sequencing platform. The bioinformatics methods were used to analyze the resistance components of p JM45-1 plasmid and clustered with the plasmid sequence NCBI (Fast Minimum Evolutin method) . Results The size of p JM45-1 plasmid was 317 156 bp. The functional annotation analysis showed that the plasmid was a joinable transfer plasmid, carrying antimicrobial resistance gene, heavy metal ion tolerance gene, toxin coding gene and transposon, as well as insertion sequence 83 resistance-related coding genes. The 45995 sequences of pMG45-1 plasmid and the full-length 94 281 sequences of drug-resistant plasmid R100 (Accession No: AP000342.1) were 99% identical to that of the conjugate plasmid F plasmid (Accession No. AP001918.1) The 8322 sequences of the 99 159 total sequences were 87% identical. p The JM45-1 plasmid is in the same cluster as the plasmid derived from Klebsiella pneumoniae and not in a cluster with plasmids derived from other Enterobacteriaceae. Conclusion Klebsiella pneumoniae JM45 p JM45-1 plasmid carrying a large number of resistant elements, is the main reason for the pan-resistant strains of the strain evolution. p JM45-1 is a conjugative transfer plasmid that transports drug-resistant genes horizontally between bacteria, resulting in the spread of resistant bacteria.