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利用 WGS 鉴定坦桑尼亚多重耐药鲍曼不动杆菌中的抗生素耐药基因并预测其抗菌药物耐药表型。

Using WGS to identify antibiotic resistance genes and predict antimicrobial resistance phenotypes in MDR Acinetobacter baumannii in Tanzania.

机构信息

Kilimanjaro Christian Medical University College, Moshi, Tanzania.

Kilimanjaro Clinical Research Institute, Moshi, Tanzania.

出版信息

J Antimicrob Chemother. 2019 Jun 1;74(6):1484-1493. doi: 10.1093/jac/dkz055.

Abstract

BACKGROUND

Reliable phenotypic antimicrobial susceptibility testing can be a challenge in clinical settings in low- and middle-income countries. WGS is a promising approach to enhance current capabilities.

AIM

To study diversity and resistance determinants and to predict and compare resistance patterns from WGS data of Acinetobacter baumannii with phenotypic results from classical microbiological testing at a tertiary care hospital in Tanzania.

METHODS AND RESULTS

MLST using Pasteur/Oxford schemes yielded eight different STs from each scheme. Of the eight, two STs were identified to be global clones 1 (n = 4) and 2 (n = 1) as per the Pasteur scheme. Resistance testing using classical microbiology determined between 50% and 92.9% resistance across all drugs. Percentage agreement between phenotypic and genotypic prediction of resistance ranged between 57.1% and 100%, with coefficient of agreement (κ) between 0.05 and 1. Seven isolates harboured mutations at significant loci (S81L in gyrA and S84L in parC). A number of novel plasmids were detected, including pKCRI-309C-1 (219000 bp) carrying 10 resistance genes, pKCRI-43-1 (34935 bp) carrying two resistance genes and pKCRI-49-1 (11681 bp) and pKCRI-28-1 (29606 bp), each carrying three resistance genes. New ampC alleles detected included ampC-69, ampC-70 and ampC-71. Global clone 1 and 2 isolates were found to harbour ISAba1 directly upstream of the ampC gene. Finally, SNP-based phylogenetic analysis of the A. baumannii isolates revealed closely related isolates in three clusters.

CONCLUSIONS

The validity of the use of WGS in the prediction of phenotypic resistance can be appreciated, but at this stage is not sufficient for it to replace conventional antimicrobial susceptibility testing in our setting.

摘要

背景

在中低收入国家的临床环境中,可靠的表型抗菌药敏试验可能是一项挑战。WGS 是增强当前能力的一种很有前途的方法。

目的

研究多样性和耐药决定因素,并预测和比较坦桑尼亚一家三级护理医院从 WGS 数据和经典微生物学测试获得的鲍曼不动杆菌的耐药模式。

方法和结果

使用 Pasteur/Oxford 方案进行 MLST,每个方案产生 8 种不同的 ST。在这 8 种 ST 中,根据 Pasteur 方案,有 2 种 ST 被鉴定为全球克隆 1(n = 4)和 2(n = 1)。使用经典微生物学进行耐药性测试,所有药物的耐药率在 50%至 92.9%之间。表型和基因型耐药预测的百分比一致性在 57.1%至 100%之间,一致性系数(κ)在 0.05 至 1 之间。7 株分离株在重要基因座(gyrA 中的 S81L 和 parC 中的 S84L)携带突变。检测到许多新型质粒,包括携带 10 个耐药基因的 pKCRI-309C-1(219000 bp)、携带 2 个耐药基因的 pKCRI-43-1(34935 bp)、携带 3 个耐药基因的 pKCRI-49-1(11681 bp)和 pKCRI-28-1(29606 bp)。检测到新的 ampC 等位基因包括 ampC-69、ampC-70 和 ampC-71。发现全球克隆 1 和 2 分离株在 ampC 基因上游直接携带 ISAba1。最后,对鲍曼不动杆菌分离株进行基于 SNP 的系统发育分析,发现三个聚类中有密切相关的分离株。

结论

可以理解 WGS 在预测表型耐药性中的有效性,但在现阶段,它还不能替代我们环境中的常规抗菌药敏试验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba1d/6524488/90533a839752/dkz055f1.jpg

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