Identification and characterization of bacterial pathogens causing bloodstream infections by DNA microarray
Identification and characterization of bacterial pathogens causing bloodstream infections by DNA microarray
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DOI：
10.1128/jcm.02291-05
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发表时间：
2006-07-01
影响因子：
9.4
通讯作者：
Krut, Oleg
中科院分区：
文献类型：
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作者：
Cleven, Berit E. E.;Palka-Santini, Maria;Krut, Oleg
血流感染可能危及生命，需要对致病病原体进行快速鉴定和抗生素敏感性测试，以促进特异性抗菌治疗。我们开发了一种原型DNA微阵列，用于鉴定和表征三种重要的引起菌血症的菌种：金黄色葡萄球菌、大肠杆菌和铜绿假单胞菌。该阵列由从重组质粒扩增的长度为200至800 bp的120个物种特异性基因探针组成。这些探针代表编码管家蛋白的基因。毒力因子和抗生素抗性决定因子。对42株临床分离株、3株参考菌株和13株阳性血培养的检测结果表明，该基因芯片对S.金黄色葡萄球菌E.大肠杆菌和铜绿假单胞菌菌株，并将它们与密切相关的革兰氏阳性和革兰氏阴性细菌菌株（也已知是菌血症的病原体）区分开来。我们发现，通过常规药敏试验确定的表型抗生素耐药性与通过与沙门氏菌杂交确定的基因型抗生素耐药性之间几乎完全相关。金黄色葡萄球菌耐药基因探针mecA（苯唑西林-甲氧西林耐药）、aacA-aphD（庆大霉素耐药）。ermA（红霉素耐药）、blaZ（青霉素耐药）和E.大肠杆菌耐药基因探针bla（TEM-106）（青霉素耐药）和aacC 2（氨基糖苷类耐药）。此外，抗生素耐药性和毒力基因探针允许在一个物种内的基因型歧视。这种新型的DNA微阵列证明了同时识别和表征血培养中的细菌的可行性，而无需事先扩增靶DNA或预先识别病原体。
Bloodstream infections are potentially life-threatening and require rapid identification and antibiotic susceptibility testing of the causative pathogen in order to facilitate specific antimicrobial therapy. We developed a prototype DNA microarray for the identification and characterization of three important bacteremia-causing species: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. The array consisted of 120 species-specific gene probes 200 to 800 bp in length that were amplified from recombinant plasmids. These probes represented genes encoding housekeeping proteins. virulence factors, and antibiotic resistance determinants. Evaluation with 42 clinical isolates, 3 reference strains, and 13 positive blood cultures revealed that the DNA microarray was highly specific in identifying S. aureus, E. coli, and P. aeruginosa strains and in discriminating them from closely, related gram-positive and gram-negative bacterial strains also known to be etiological agents of bacteremia. We found a nearly perfect correlation between phenotypic antibiotic resistance determined by conventional susceptibility testing and genotypic antibiotic resistance by hybridization to the S. aureus resistance gene probes mecA (oxacillin-methicillin resistance), aacA-aphD (gentamicin resistance). ermA (erythromycin resistance), and blaZ (penicillin resistance) and the E. coli resistance gene probes bla(TEM-106) (penicillin resistance) and aacC2 (aminoglycoside resistance). Furthermore, antibiotic resistance and virulence gene probes permitted genotypic discrimination within a species. This novel DNA microarray demonstrates the feasibility of simultaneously identifying and characterizing bacteria in blood cultures without prior amplification of target DNA or preidentification of the pathogen.