Missoni Emilija Mlinarić, Kalenić Smilja, Vukelić Milan, De Syo Drago, Belicza Mladen, Kern Josipa, Babić Verica Vazić
Odsjek za klinicku mikologiju, Hrvatski zavod za javno zdravstvo, Zagreb.
Acta Med Croatica. 2006;60(1):43-50.
The aim was to assess the incidence of isolation of individual fungal species and interpret the meaning of fungal isolates from foot ulcers of 509 diabetic outpatients using mycologic and histopathologic methods. Another aim was to explore risk factors for the development of fungal infections in foot ulcer.
Fungus isolation was made on selective media and their identification by standard mycologic methods. Histopathologic diagnosis of fungal ulcer infections was made on PAS-stained histopathologic preparations and imprint preparations (PAS and Papanicolaou staining) of foot wound biopsy specimens.
Fungal and mixed foot ulcer infections were found in 14.9% of diabetic patients. In 33.8% of patients, these infections were confirmed by a finding of fungal elements in histopathologic preparations of ulcer biopsy specimens, as follows: in 16.9% of patients, by finding fungal elements in imprint preparations of ulcer biopsy specimens and by isolation fungus from the swab of the same ulcer; in 2.3% by fungus isolation from ulcer biopsy specimens; in 36.9% by fungus isolation from ulcer swabs in pure culture and/or in a large number of colonies and/or from several ulcers on the foot of the same patient. More than 89% of patients had a single foot ulcer with fungal or mixed infection, big toe and the plantar-metatarsal region in one foot or both feet being the most common sites of ulcer. Fifteen species from the genera Candida, Cryptococcus, Trichosporon and Rhodotorula were the causative agents of fungal and mixed foot ulcer infections. C. parapsilosis (in 61.5% of patients), and C. albicans and C. tropicalis (in 10.8% of patients each) were the most common causes of these infections. The presence of yeasts and/or dermatophytes in the toe web of the same or other foot, or of both feet, did not influence the incidence of fungal and mixed foot ulcer infections. Patient sex and age, type and length of diabetes, or clinical picture of diabetic foot did not affect it either. In IDDM patients, the risk factor for the development of fungal and mixed foot ulcer infections was ulcer infection lasting for more than 13 weeks, whereas in NIDDM patients the length of ulcer infection did not contribute to the incidence of fungal and mixed foot ulcer infection.
Our results and other reports suggest that Candida species are the most common fungal isolates (between 93.2% and 100% of all fungal isolates) from diabetic foot ulcer, with C. parapsilosis being the most common causative agent of fungal and mixed infection. From diabetic foot ulcer, bacterial isolation was 5 times as common as that of yeasts (327 vs. 65 patients). Nevertheless, this investigation showed fungal isolates, originating not only from a primarily sterile ulcer sample (biopsy specimen) but also from foot ulcer swabs to be the causative agents (not ulcer colonizers or contaminants) of the foot ulcer infection. The pathogen c effect of yeasts in foot ulcer is indicated by the severity of clinical finding, chronic course of infection, and infection progression despite antibiotic therapy. Equally indicative are microbiologic diagnostic parameters (isolation in pure culture, and/or isolation in a large number of colonies, and/or isolation from several ulcers in the foot of the same patient).
In diabetic patients at highest risk of developing fungal and mixed foot ulcer infections (IDDM patients with ulcer infection persisting for more than 13 weeks, and NIDDM patients with the clinical picture of deep ulcer and abscess in the plantar region, irrespective of the duration of ulcer infection), routine bacteriologic diagnosis should be supplemented with targeted mycologic and histopathologic methods.
本研究旨在评估509例糖尿病门诊患者足部溃疡中分离出的各真菌种类的发生率,并运用真菌学和组织病理学方法阐释真菌分离株的意义。另一个目的是探究足部溃疡真菌感染发生的危险因素。
在选择性培养基上进行真菌分离,并通过标准真菌学方法进行鉴定。对足部伤口活检标本的PAS染色组织病理学切片及印片标本(PAS和巴氏染色)进行真菌溃疡感染的组织病理学诊断。
14.9%的糖尿病患者存在真菌及混合性足部溃疡感染。33.8%的患者经溃疡活检标本的组织病理学切片中发现真菌成分得以确诊,具体情况如下:16.9%的患者在溃疡活检标本印片中发现真菌成分且从同一溃疡拭子中分离出真菌;2.3%的患者从溃疡活检标本中分离出真菌;36.9%的患者从溃疡拭子中分离出纯培养真菌和/或大量菌落和/或从同一患者足部多个溃疡中分离出真菌。超过89%的患者有单个足部溃疡伴真菌或混合感染,单足或双足的大脚趾及跖趾区域是最常见的溃疡部位。念珠菌属、隐球菌属、丝孢酵母属和红酵母属的15个菌种是真菌及混合性足部溃疡感染的病原体。近平滑念珠菌(61.5%的患者)、白色念珠菌和热带念珠菌(各占10.8%的患者)是这些感染最常见的病因。同一或另一只脚或双脚的趾间有酵母菌和/或皮肤癣菌存在,并不影响真菌及混合性足部溃疡感染的发生率。患者的性别、年龄、糖尿病类型及病程,或糖尿病足的临床表现也未对其产生影响。在胰岛素依赖型糖尿病(IDDM)患者中,真菌及混合性足部溃疡感染发生的危险因素是溃疡感染持续超过13周,而在非胰岛素依赖型糖尿病(NIDDM)患者中,溃疡感染时长对真菌及混合性足部溃疡感染的发生率并无影响。
我们的研究结果及其他报告表明,念珠菌属是糖尿病足部溃疡中最常见的真菌分离株(占所有真菌分离株的93.2%至100%),近平滑念珠菌是真菌及混合感染最常见的病原体。糖尿病足部溃疡中细菌分离的发生率是酵母菌的5倍(327例对65例患者)。然而,本研究显示,真菌分离株不仅来源于原本无菌的溃疡样本(活检标本),还来源于足部溃疡拭子,是足部溃疡感染的病原体(而非溃疡定植菌或污染物)。临床症状的严重程度、感染的慢性病程以及尽管使用抗生素治疗但感染仍进展,均表明酵母菌在足部溃疡中的致病作用。同样具有指示性的是微生物学诊断参数(纯培养分离、和/或大量菌落分离、和/或从同一患者足部多个溃疡中分离)。
对于发生真菌及混合性足部溃疡感染风险最高的糖尿病患者(溃疡感染持续超过13周的IDDM患者,以及足底区域有深部溃疡和脓肿临床表现的NIDDM患者,无论溃疡感染时长),常规细菌学诊断应辅以针对性的真菌学和组织病理学方法。