背景:弧菌是在地表水中发现的自生细菌种群,与各种危及生命的肠外疾病有关,特别是在患有潜在疾病和伤口感染的人群中。目前,关于这些物种的毒力和抗性基因的突变多样性,存在非常小的信息。这项研究评估了核酸内切酶的变异以及弧菌分离株的毒力和抗性基因的突变多样性,携带毒力相关基因(vcgCPI),二氢蝶呤合酶1型和II型基因(Sul1和11),(aadA)氨基糖苷(3\'\')(9)腺苷酸转移酶基因,(aac(3)-IIa,(aacC2)a,氨基糖苷N(3)-乙酰转移酶III,和(strA)氨基糖苷3'-磷酸转移酶抗性基因。
方法:使用分子生物学技术的组合,生物信息学工具,和序列分析。
结果:我们的结果揭示了在核苷酸位置(密码子)73-75(A→G)和300-302(N→S)处,创伤弧菌(vcgCPI)的毒力决定子的各种核苷酸变异。弧菌属的氨基糖苷类抗性基因(aadA)描述了482位的核苷酸差异(A→G),而氨基糖苷类耐药基因(sul1和11)显示两个核苷酸多态性可变区(102和140)。氨基酸差异存在于位置140的核苷酸多态性(A→E)。限制酶HinP1I产生的条带模式,MwoI,和StyD4I显示出显著的变化。此外,蛋白质二氢蝶呤合酶1型和II型基因(Sul1和11)的限制酶消化显着不同,而酶DpnI和Hinf1表示没有显著差异。与来自GenBank的参考分离株相比,限制酶NlaIV没有显示条带。然而,抗性决定簇显示显著的点核苷酸突变,不会产生任何具有不同多态性区域的氨基酸变化,正如限制摘要中所揭示的那样。
结论:描述的毒力和抗性决定子具有与致病基因组学研究相关的特定多态性位点,药物基因组学,和控制这些与水相关的菌株。
BACKGROUND: Vibrio species are among the autochthonous bacterial populations found in surface waters and associated with various life-threatening extraintestinal diseases, especially in human populations with underlying illnesses and wound infections. Presently, very diminutive information exists regarding these species\' mutational diversity of virulence and resistance genes. This study evaluated variations in endonucleases and mutational diversity of the virulence and resistance genes of Vibrio isolates, harboring virulence-correlated gene (vcgCPI), dihydropteroate synthase type 1 and type II genes (Sul 1 and 11), (aadA) aminoglycoside (3\'\') (9) adenylyltransferase gene, (aac(3)-IIa, (aacC2)a, aminoglycoside N(3)-acetyltransferase III, and (strA) aminoglycoside 3\'-phosphotransferase resistance genes.
METHODS: Using combinations of molecular biology techniques, bioinformatics tools, and sequence analysis.
RESULTS: Our result revealed various nucleotide variations in virulence determinants of V. vulnificus (vcgCPI) at nucleotide positions (codon) 73-75 (A → G) and 300-302 (N → S). The aminoglycosides resistance gene (aadA) of Vibrio species depicts a nucleotide difference at position 482 (A → G), while the aminoglycosides resistance gene (sul 1 and 11) showed two variable regions of nucleotide polymorphism (102 and 140). The amino acid differences exist with the nucleotide polymorphism at position 140 (A → E). The banding patterns produced by the restriction enzymes HinP1I, MwoI, and StyD4I showed significant variations. Also, the restriction enzyme digestion of protein dihydropteroate synthase type 1 and type II genes (Sul 1 and 11) differed significantly, while enzymes DpnI and Hinf1 indicate no significant differences. The restriction enzyme NlaIV showed no band compared to reference isolates from the GenBank. However, the resistant determinants show significant point nucleotide mutation, which does not produce any amino acid change with diverse polymorphic regions, as revealed in the restriction digest profile.
CONCLUSIONS: The described virulence and resistance determinants possess specific polymorphic locus relevant to pathogenomics studies, pharmacogenomic, and control of such water-associated strains.