在乌干达西南部地区从地表水中回收的人类致病性弧菌物种中,毒力(vcgCPI/vcgCPE)和抗性决定簇(aac(3)-IIa、(aacC2、strA、Sul 1和11)的多态性和突变多样性。

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal, genetic engineering & biotechnology Pub Date : 2023-10-06 DOI:10.1186/s43141-023-00554-1
Hope Onohuean, Uchechukwu U Nwodo
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引用次数: 0

摘要

背景:弧菌是地表水中发现的本地细菌种群之一,与各种危及生命的肠道外疾病有关,尤其是在患有潜在疾病和伤口感染的人群中。目前,关于这些物种毒力和抗性基因的突变多样性的信息非常少。本研究评估了弧菌分离株的核酸内切酶变异以及毒力和抗性基因的突变多样性,这些分离株携带毒力相关基因(vcgCPI)、1型和II型二氢翼酸合成酶基因(Sul 1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:结合分子生物学技术、生物信息学工具和序列分析。结果:我们的结果揭示了创伤弧菌(vcgCPI)毒力决定簇在核苷酸位置(密码子)73-75(A → G) 和300-302(N → S) 。弧菌属物种的氨基糖苷类抗性基因(aadA)在482位(a → G) ,而氨基糖苷类抗性基因(sul 1和11)显示出两个核苷酸多态性可变区(102和140)。140位核苷酸多态性存在氨基酸差异(A → E) 。限制性内切酶HinP1I、MwoI和StyD4I产生的条带模式显示出显著的变化。此外,蛋白二氢翼酸合成酶1型和II型基因(Sul 1和11)的限制性内切酶消化显著不同,而酶DpnI和Hinf1没有显著差异。与来自GenBank的参考分离株相比,限制性内切酶NlaIV没有显示出条带。然而,如限制性消化图谱所示,抗性决定簇显示出显著的点核苷酸突变,这不会产生任何具有不同多态性区域的氨基酸变化。结论:所描述的毒力和抗性决定簇具有与病原学研究、药物基因组学和水相关菌株控制相关的特异性多态位点。
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Polymorphism and mutational diversity of virulence (vcgCPI/vcgCPE) and resistance determinants (aac(3)-IIa, (aacC2, strA, Sul 1, and 11) among human pathogenic Vibrio species recovered from surface waters in South-Western districts of Uganda.

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.

Conclusion: The described virulence and resistance determinants possess specific polymorphic locus relevant to pathogenomics studies, pharmacogenomic, and control of such water-associated strains.

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