Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University, Aalborg, Denmark.
Wound Repair Regen. 2010 Jan-Feb;18(1):38-49. doi: 10.1111/j.1524-475X.2009.00561.x.
The bacterial microbiota plays an important role in the prolonged healing of chronic venous leg ulcers. The present study compared the bacterial diversity within ulcer material from 14 skin graft operations of chronic venous leg ulcers using culture-based methods and molecular biological methods, such as 16S rRNA gene sequencing, fingerprinting, quantitative polymerase chain reaction, and fluorescence in situ hybridization. Each wound contained an average of 5.4 species but the actual species varied between wounds. The diversity determined by culture-based methods and the molecular biological methods was different. All the wounds contained Staphylococcus aureus, whereas Pseudomonas aeruginosa was in six out of 14 wounds. Molecular methods detected anaerobic pathogens in four ulcers that were not detected with anaerobic culture methods. Quantitative polymerase chain reaction was used to compare the abundance of S. aureus and P. aeruginosa at different locations in the ulcers and their numbers varied greatly between samples taken at different locations in the same ulcer. This should be considered when ulcers are investigated in routine clinical care. The differences between the results obtained with culture-based and molecular-based approaches demonstrate that the use of one approach alone is not able to identify all of the bacteria present in the wounds.
细菌菌群在慢性静脉性腿部溃疡的长期愈合中起着重要作用。本研究比较了 14 例慢性静脉性腿部溃疡皮肤移植手术中溃疡材料的细菌多样性，使用了基于培养的方法和分子生物学方法，如 16S rRNA 基因测序、指纹图谱、定量聚合酶链反应和荧光原位杂交。每个伤口平均含有 5.4 种，但实际物种在伤口之间存在差异。基于培养的方法和分子生物学方法确定的多样性不同。所有伤口均含有金黄色葡萄球菌，而 14 个伤口中有 6 个含有铜绿假单胞菌。分子方法检测到 4 个溃疡中有厌氧病原体，而厌氧培养方法未检测到。定量聚合酶链反应用于比较溃疡不同部位金黄色葡萄球菌和铜绿假单胞菌的丰度，其数量在同一溃疡不同部位采集的样本之间差异很大。在常规临床护理中对溃疡进行研究时应考虑到这一点。基于培养和基于分子的方法的结果差异表明，单独使用一种方法无法识别伤口中存在的所有细菌。
