Mixed-genome microarrays reveal multiple serotype and lineage-specific differences among strains of Listeria monocytogenes
Mixed-genome microarrays reveal multiple serotype and lineage-specific differences among strains of Listeria monocytogenes
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DOI：
10.1128/jcm.41.2.632-639.2003
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发表时间：
2003-02-01
影响因子：
9.4
通讯作者：
Besser, TE
中科院分区：
文献类型：
--
作者：
Call, DR;Borucki, MK;Besser, TE
流行病学研究和推定的毒力基因的分析表明，单核细胞增生李斯特菌已分化成几个系统发育司。我们假设，许多基因，影响生态位特异性健身和毒力，并确定这些差异可能会提供新的机会，检测，治疗和控制这种重要的病原体发生了类似的分歧。为了进一步探讨这个问题，我们开发了一种由取自10株L.单核细胞增多症然后，我们将来自50种不同菌株的基因组DNA与重复阵列杂交，并分析了所得的杂交模式。一个简单的欧几里德距离度量允许重建先前描述的血清型之间的遗传关系，并且仅需要四个微阵列探针来区分最重要的血清型（1/2a，1/2b，1/2c和4）。我们计算了基因芯片数据的连锁平衡指数，证实了L。单核细胞增多症具有强克隆性群体结构（I-A = 3.85）。从文库中检索到29个信息探针并进行测序。这些包括与修复紫外线损伤的DNA，耐盐性，生物膜形成，重金属运输，亚铁运输和磷壁酸合成相关的基因。几个膜结合脂蛋白和一个interalin被确定，加上三个噬菌体序列和六个序列与未知功能。总的来说，这些数据证实了许多基因在L.单核细胞增多症此外，这些结果证明了混合基因组微阵列作为衍生生物学有用信息以及用于识别和筛选临床重要微生物的遗传标记的工具的价值。
Epidemiological studies and analysis of putative virulence genes have shown that Listeria monocytogenes has diverged into several phylogenetic divisions. We hypothesize that similar divergence has occurred for many genes that influence niche-specific fitness and virulence and that identifying these differences may offer new opportunities for the detection, treatment, and control of this important pathogen. To explore this issue further, we developed a microarray composed of fragmented DNA taken from 10 strains of L. monocytogenes. We then hybridized genomic DNA from 50 different strains to replicate arrays and analyzed the resulting hybridization patterns. A simple Euclidean distance metric permitted the reconstruction of previously described genetic relationships between serotypes, and only four microarray probes were needed to discriminate between the most important serotypes (1/2a, 1/2b, 1/2c, and 4). We calculated an index of linkage equilibrium from the microarray data and confirmed that L. monocytogenes has a strongly clonal population structure (I-A = 3.85). Twenty-nine informative probes were retrieved from the library and sequenced. These included genes associated with repairing UV-damaged DNA, salt tolerance, biofilm formation, heavy metal transport, ferrous iron transport, and teichoic acid synthesis. Several membrane-bound lipoproteins and one internalin were identified, plus three phage sequences and six sequences with unknown function. Collectively, these data confirm that many genes have diverged between lineages of L. monocytogenes. Furthermore, these results demonstrate the value of mixed-genome microarrays as a tool for deriving biologically useful information and for identifying and screening genetic markers for clinically important microbes.