Bosch Thijs, Verkade Erwin, van Luit Martijn, Pot Bruno, Vauterin Paul, Burggrave Ronald, Savelkoul Paul, Kluytmans Jan, Schouls Leo
Laboratory for Infectious Diseases and Screening, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.
PLoS One. 2013 Jun 21;8(6):e66493. doi: 10.1371/journal.pone.0066493. Print 2013.
After its emergence in 2003, a livestock-associated (LA-)MRSA clade (CC398) has caused an impressive increase in the number of isolates submitted for the Dutch national MRSA surveillance and now comprises 40% of all isolates. The currently used molecular typing techniques have limited discriminatory power for this MRSA clade, which hampers studies on the origin and transmission routes. Recently, a new molecular analysis technique named whole genome mapping was introduced. This method creates high-resolution, ordered whole genome restriction maps that may have potential for strain typing. In this study, we assessed and validated the capability of whole genome mapping to differentiate LA-MRSA isolates. Multiple validation experiments showed that whole genome mapping produced highly reproducible results. Assessment of the technique on two well-documented MRSA outbreaks showed that whole genome mapping was able to confirm one outbreak, but revealed major differences between the maps of a second, indicating that not all isolates belonged to this outbreak. Whole genome mapping of LA-MRSA isolates that were epidemiologically unlinked provided a much higher discriminatory power than spa-typing or MLVA. In contrast, maps created from LA-MRSA isolates obtained during a proven LA-MRSA outbreak were nearly indistinguishable showing that transmission of LA-MRSA can be detected by whole genome mapping. Finally, whole genome maps of LA-MRSA isolates originating from two unrelated veterinarians and their household members showed that veterinarians may carry and transmit different LA-MRSA strains at the same time. No such conclusions could be drawn based spa-typing and MLVA. Although PFGE seems to be suitable for molecular typing of LA-MRSA, WGM provides a much higher discriminatory power. Furthermore, whole genome mapping can provide a comparison with other maps within 2 days after the bacterial culture is received, making it suitable to investigate transmission events and outbreaks caused by LA-MRSA.
2003年出现后,一种与家畜相关的耐甲氧西林金黄色葡萄球菌(LA-MRSA)进化枝(CC398)使得提交给荷兰全国耐甲氧西林金黄色葡萄球菌监测的分离株数量显著增加,目前占所有分离株的40%。目前使用的分子分型技术对这种耐甲氧西林金黄色葡萄球菌进化枝的鉴别能力有限,这妨碍了对其起源和传播途径的研究。最近,引入了一种名为全基因组图谱的新分子分析技术。该方法可创建高分辨率、有序的全基因组限制性图谱,可能具有菌株分型的潜力。在本研究中,我们评估并验证了全基因组图谱区分LA-MRSA分离株的能力。多项验证实验表明,全基因组图谱产生了高度可重复的结果。对两次记录良好的耐甲氧西林金黄色葡萄球菌暴发进行该技术评估表明,全基因组图谱能够确认一次暴发,但揭示了第二次暴发图谱之间的重大差异,表明并非所有分离株都属于此次暴发。对流行病学上无关联的LA-MRSA分离株进行全基因组图谱分析提供了比spa分型或多位点可变数目串联重复序列分析(MLVA)更高的鉴别能力。相比之下,在一次已证实的LA-MRSA暴发期间获得的LA-MRSA分离株所创建的图谱几乎无法区分,表明通过全基因组图谱可以检测到LA-MRSA的传播。最后,来自两名不相关兽医及其家庭成员的LA-MRSA分离株的全基因组图谱表明,兽医可能同时携带和传播不同的LA-MRSA菌株。基于spa分型和MLVA无法得出这样的结论。虽然脉冲场凝胶电泳(PFGE)似乎适用于LA-MRSA的分子分型,但全基因组图谱提供了更高的鉴别能力。此外,全基因组图谱可以在收到细菌培养物后2天内与其他图谱进行比较,使其适合于调查由LA-MRSA引起的传播事件和暴发。