Nova Research Inc., Alexandria, VA, 22308, USA.
Center for Biomolecular Science & Engineering, US Naval Research Laboratory, Washington, DC, USA.
BMC Res Notes. 2024 Mar 12;17(1):70. doi: 10.1186/s13104-024-06717-8.
In this study, we sought to determine the types and prevalence of antimicrobial resistance determinants (ARDs) in Burkholderia spp. strains using the Antimicrobial Resistance Determinant Microarray (ARDM).
Whole genome amplicons from 22 B. mallei (BM) and 37 B. pseudomallei (BP) isolates were tested for > 500 ARDs using ARDM v.3.1. ARDM detected the following Burkholderia spp.-derived genes, aac(6), bla, blaA, penA-BP, and qacE, in both BM and BP while bla, macB, bla and penA-BC were observed in BP only. The method of denaturing template for whole genome amplification greatly affected the numbers and types of genes detected by the ARDM. Bla was detected in nearly a third of BM and BP amplicons derived from thermally, but not chemically denatured templates. Bla results were confirmed by PCR, with 81% concordance between methods. Sequences from 414-nt PCR amplicons (13 preparations) were 100% identical to the Klebsiella pneumoniae reference gene. Although bla sequences have been observed in B. glumae, B. cepacia, and other undefined Burkholderia strains, this is the first report of such sequences in BM/BP/B. thailandensis (BT) clade. These results highlight the importance of sample preparation in achieving adequate genome coverage in methods requiring untargeted amplification before analysis.
本研究使用抗菌药物耐药决定因子微阵列（ARDM）检测伯克霍尔德菌属菌株中抗菌药物耐药决定因子（ARDs）的类型和流行率。
使用 ARDM v.3.1 对 22 株鼻疽伯克霍尔德菌（BM）和 37 株类鼻疽伯克霍尔德菌（BP）的全基因组扩增子进行了>500 种 ARD 的检测。ARDM 检测到 BM 和 BP 中均存在以下伯克霍尔德菌属衍生基因：aac(6)、bla、blaA、penA-BP 和 qacE，而 bla、macB、bla 和 penA-BC 仅在 BP 中观察到。全基因组扩增模板的变性方法极大地影响了 ARDM 检测到的基因数量和类型。bla 在近三分之一的来源于热变性而不是化学变性模板的 BM 和 BP 扩增子中被检测到。bla 结果通过 PCR 得到证实，两种方法的一致性为 81%。来自 414-nt PCR 扩增子（13 个制剂）的序列与肺炎克雷伯菌参考基因完全一致。尽管 bla 序列已在稻黄伯克霍尔德菌、洋葱伯克霍尔德菌和其他未定义的伯克霍尔德菌菌株中观察到，但这是首次在 BM/BP/B. thailandensis（BT）进化枝中报告此类序列。这些结果强调了在需要在分析前进行非靶向扩增的方法中，样品制备对于实现充分的基因组覆盖的重要性。
