Rapid molecular detection of macrolide resistance
Rapid molecular detection of macrolide resistance
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DOI：
10.1186/s12879-019-3762-4
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发表时间：
2019-02-12
影响因子：
3.7
通讯作者：
Bracht, John R.
中科院分区：
文献类型：
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作者：
Nelson, Megan M.;Waldron, Christopher L.;Bracht, John R.
背景：新出现的抗菌素耐药性是对人类健康的重大威胁。然而，快速诊断抗菌素耐药性的方法通常需要基于多天培养的检测。大环内酯类抗生素外排基因MEF(A)对红霉素和阿奇霉素具有抗药性，可以在多种细菌之间横向转移。方法：采用重组酶聚合酶分析法(RPA)从未经核酸纯化的裂解产物中检测抗菌素基因MEF(A)。为了验证这些结果，我们进行了肉汤稀释法来评估红霉素和氨苄西林(阴性对照)的耐药性。结果：我们在7-10分钟内验证了化脓性链球菌、肺炎链球菌、唾液链球菌和粪肠球菌细菌裂解物中MEF(A)的检测。我们表明，MEF(A)的检测准确地预测了传统培养方法评估的真实抗菌素耐药性，并且该检测对高水平的掺入非特异性核酸污染是稳健的。该检测方法不受不同MEF(A)基因序列中单核苷酸多态性的影响，加强了其作为一种可靠诊断工具的实用性。结论这一发现为在临床环境中实施快速基因组诊断打开了大门，同时为研究人员提供了一种快速、经济有效的工具来追踪病原菌和共生菌株的抗生素耐药性。
Background: Emerging antimicrobial resistance is a significant threat to human health. However, methods for rapidly diagnosing antimicrobial resistance generally require multi-day culture-based assays. Macrolide efflux gene A, mef(A), provides resistance against erythromycin and azithromycin and is known to be laterally transferred among a wide range of bacterial species.Methods: We use Recombinase Polymerase Assay (RPA) to detect the antimicrobial resistance gene mef(A) from raw lysates without nucleic acid purification. To validate these results we performed broth dilution assays to assess antimicrobial resistance to erythromycin and ampicillin (a negative control).Results: We validate the detection of mef(A) in raw lysates of Streptococcus pyogenes, S. pneumoniae, S. salivarius, and Enterococcus faecium bacterial lysates within 7-10 min of assay time. We show that detection of mef(A) accurately predicts real antimicrobial resistance assessed by traditional culture methods, and that the assay is robust to high levels of spiked-in non-specific nucleic acid contaminant. The assay was unaffected by single-nucleotide polymorphisms within divergent mef(A) gene sequences, strengthening its utility as a robust diagnostic tool.Conclusions This finding opens the door to implementation of rapid genomic diagnostics in a clinical setting, while providing researchers a rapid, cost-effective tool to track antibiotic resistance in both pathogens and commensal strains.