回顾性分析慢性难愈创面患者病原菌分布特征和耐药情况，以指导临床制定个性化方案并合理使用抗菌药物。

对2020年1月至2025年1月清华大学第一附属医院皮肤科门诊、皮肤科病房及血管外科病房收治的124例慢性难愈创面患者的临床资料进行回顾性研究。用无菌拭子取患者皮肤溃疡创面分泌物或刮取坏死组织送实验室进行细菌培养。菌种鉴定采用法国生物梅里埃公司VITEK MS检测系统VITEK-2鉴定卡，体外药敏试验采用配套VITEK-2药敏卡及K-B纸片（英国Oxoid公司）扩散法。依据美国临床实验室标准化协会（CLSI）标准进行试验操作及结果判读。应用二元Logistic回归分析创面病原菌培养阳性的独立危险因素。

共分离出病原菌92株，分离率为74.2%，其中革兰阳性球菌主要为金黄色葡萄球菌（27株、29.3%）和凝固酶阴性葡萄球菌（23株、25%）；革兰阴性杆菌主要为铜绿假单胞菌（9株、9.8%）、大肠埃希菌（4株、4.3%）和奇异变形杆菌（4株、4.3%）。慢性难愈创面的病因居前3位的分别为下肢动脉血管性溃疡（43例、34.7%）、感染性溃疡（38例、30.6%）和术后伤口愈合不良（15例、12.1%）。感染性溃疡患者金黄色葡萄球菌的检出率显著高于下肢动脉血管病变患者（χ²=6.618、P=0.014）。≥ 60岁患者创面病原菌检出率较< 60岁患者显著增高（χ²=5.236、P=0.022）。Logistic回归分析显示，糖尿病是创面病原菌培养阳性的独立危险因素（OR=2.620、95%CI：1.013～6.777、P=0.047）。糖尿病患者创面病原菌检出率显著高于无糖尿病患者（χ²=7.079、P=0.008），糖尿病患者其他革兰阴性杆菌的阳性检出率显著高于无糖尿病患者（χ²=3.932、P=0.047），糖尿病病程≥15年患者其他革兰阴性杆菌检出率显著高于病程< 15年患者（χ²=5.013、P=0.025），差异均有统计学意义。头颈部金黄色葡萄球菌检出率显著高于躯干部（χ²=8.531、P=0.003）和四肢（χ²=11.738、P=0.001）。耐甲氧西林金黄色葡萄球菌（5株、5.4%）和耐甲氧西林凝固酶阴性葡萄球菌（15株、16.3%）均对万古霉素、替加环素和利奈唑胺敏感。9株铜绿假单胞菌（9.8%）对头孢吡肟、美罗培南、阿米卡星、左旋氧氟沙星、环丙沙星和黏菌素均敏感，1株（11.1%）对亚胺培南耐药。4株大肠埃希菌（4.3%）中对阿米卡星、左旋氧氟沙星和磺胺甲恶唑/甲氧苄啶各出现1株（25%）耐药。4株奇异变形杆菌（4.3%）中出现3株（75%）对替加环素耐药，各出现2株（50%）对头孢呋肟及磺胺甲恶唑/甲氧苄啶耐药。

感染与慢性难愈创面的发生关系密切，糖尿病患者及老年患者是高发人群，创面细菌种类繁多，分布有临床特点，个体化合理选择抗菌药物可以有效控制感染，促进创面愈合，预防和延缓耐药菌的产生和传播。

To investigate the distribution characteristics and drug resistance of pathogenic bacteria in patients with chronic refractory wounds, and to guide the formulation of personalized antimicrobial treatment plans and promote rational use of antibacterial drug in clinical practice.

A retrospective study was conducted on the clinical data of 124 patients with chronic refractory wounds admitted to the Dermatology Outpatient Department, Dermatology Inpatient Unit and Vascular Surgery Inpatient Unit of the First Affiliated Hospital of Tsinghua University from January 2020 to January 2025. Wound secretions or necrotic tissues were collected by sterile swabs and sent to laboratory for bacterial culture. Species identification was performed by VITEK MS microbial identification system (bioMérieux, France) with VITEK-2 identification cards. In vitro antimicrobial susceptibility testing employed matching VITEK-2 susceptibility cards and the Kirby-Bauer disk diffusion method (Oxoid, UK). All experimental procedures and result interpretations were adhered to guidelines established by the Clinical and Laboratory Standards Institute (CLSI). The independent risk factors for positive pathogen culture in wound were analyzed by binary Logistic regression.

Total of 92 strains of pathogenic bacteria were isolated, with the isolation rate of 74.2%; Gram-positive cocci primarily included Staphylococcus aureus (27 strains, 29.3%) and coagulase-negative Staphylococci (23 strains, 25%); Gram-negative bacilli mainly comprised Pseudomonas aeruginosa (9 strains, 9.8%), Escherichia coli (4 strains, 4.3%) and Proteus mirabilis (4 strains, 4.3%). The top three etiological factors for chronic refractory wounds were lower extremity arterial vascular ulcers (43 cases, 34.7%), infectious ulcers (38 cases, 30.6%) and postoperative poor wound healing (15 cases, 12.1%). The detection rate of Staphylococcus aureus was significantly higher in patients with infectious ulcer group compared with patients of lower extremity arterial disease (χ²=6.618, P=0.014). Wound pathogen detection rates were significantly elevated in patients ≥ 60 years old compared with those < 60 years old (χ²=5.236, P=0.022). Logistic regression analysis showed that diabetes mellitus was an independent risk factor for positive wound pathogen culture (OR=2.620, 95%CI: 1.013-6.777, P=0.047). The detection rate of wound pathogen was significantly higher in patients with diabetes compared with patients without diabetes (χ²=7.079, P=0.008), the positive detection rate of other Gram-negative bacilli was also significantly higher in patients with diabetic (χ²=3.932, P=0.047). In patients with type 2 diabetes mellitus duration ≥ 15 years, detection rates of other Gram-negative bacilli were significantly higher than those with duration < 15 years (χ²=5.013, P=0.025), all with significant differences. The detection rate of Staphylococcus aureus in head and neck region was significantly higher than that in trunk (χ²=8.531, P=0.003) and limbs (χ²=11.738, P=0.001). Methicillin-resistant Staphylococcus aureus accounted for 5 strains (5.4%) and methicillin-resistant coagulase-negative Staphylococci accounted for 15 strains (16.3%), all were sensitive to vancomycin, tigecycline and linezolid. Pseudomonas aeruginosa (9 strains, 9.8%) demonstrated universal susceptibility to cefepime, meropenem, amikacin, levofloxacin, ciprofloxacin and colistin, while 1 strain (11.1%) exhibited resistance to imipenem. Among Escherichia coli isolates (4 strains, 4.3%), resistance was observed in 1 strain (25%) to amikacin, levofloxacin and sulfamethoxazole/trimethoprim, respectively. Proteus mirabilis (4 strains, 4.3%) showed resistance to tigecycline in 3 strains (75%) and 2 strains (50%) to cefuroxime and sulfamethoxazole/trimethoprim, respectively.

Infection is closely associated with the development of chronic refractory wounds, with diabetic and elderly patients being particularly high-risk populations. Wound bacteria exhibit diverse species and characteristic clinical distribution patterns. Rational individualized selection of antimicrobial agents can effectively control infection, promote wound healing, and prevent or delay the emergence and spread of antibiotic-resistant bacteria.