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[从有阴道不适症状女性的阴道及宫内节育器尾丝样本中分离出的念珠菌属的体外生物膜形成及其与抗真菌耐药性的关系]

[In vitro biofilm formation and relationship with antifungal resistance of Candida spp. isolated from vaginal and intrauterine device string samples of women with vaginal complaints].

作者信息

Calışkan Seyda, Keçeli Özcan Sema, Cınar Selvi, Corakçı Aydın, Calışkan Eray

机构信息

Kocaeli University Faculty of Medicine, Department of Medical Microbiology, Kocaeli, Turkey.

出版信息

Mikrobiyol Bul. 2011 Oct;45(4):697-706.

PMID:22090300
Abstract

Intrauterin device (IUD) application is a widely used effective, safe and economic method for family planning. However IUD use may cause certain changes in vaginal ecosystem and may disturb microflora leading to increased colonization of various opportunistic pathogen microorganisms. The aims of this study were (i) to detect the biofilm production characteristics of Candida spp. isolated from vaginal and IUD string samples of women with IUDs, and (ii) to investigate the relationship between biofilm production and antifungal resistance. A total of 250 women (mean age: 34.4 ± 7.6 years) admitted to gynecology outpatient clinics with vaginal symptoms (discharge and itching) were included in the study. The patients have been implanted CuT380a type IUDs for a mean duration of 59.8 ± 42.4 months. Without removing IUD, string samples were obtained by cutting and simultaneous vaginal swab samples were also collected. Isolated Candida spp. were identified by conventional methods and API 20C AUX (BioMerieux, Fransa) system. Minimal inhibitory concentrations (MIC) of fluconazole, itraconazole and amphotericin B were determined by broth microdilution method according to the CLSI guidelines. Biofilm formation was evaluated by crystal violet staining and XTT-reduction assays, and the isolates which yielded positive results in both of the methods were accepted as biofilm-producers. In the study, Candida spp. were isolated from 33.2% (83/250) of the vaginal and 34% (85/250) of the IUD string samples, C.albicans being the most frequently detected species (54 and 66 strains for the samples, respectively). The total in vitro biofilm formation rate was 25% (21/83) for vaginal isolates and 44.7% (38/85) for IUD string isolates. Biofilm formation rate of vaginal C.albicans isolates was significantly lower than vaginal non-albicans Candida spp. (14.8% and 44.8%, respectively; p= 0.003). Biofilm formation rate of C.albicans strains isolated from vaginal and IUD string samples were found as 14.8% (8/54) and 45.5% (30/66), with a statistically significant importance (p< 0.001). However, no statistically significant difference was detected for biofilm formation rates of non-albicans Candida spp. when sample types were considered [44.8% (13/29) and 42.1% (8/19), respectively; p> 0.05]. Fluconazole resistance was significantly higher in biofilm-producing vaginal Candida spp. than those of nonproducers (52.4% vs. 16.1%; p= 0.001), however, itraconazole resistance was found similar in biofilmproducer and non-producer isolates (47.6% vs. 32.3%; p> 0.05). Resistance rates for both fluconazole and itraconazole were higher in biofilm-producers (39.5% and 52.6%, respectively), than those of non-producers (10.6% and 29.8%, respectively), representing a statistical significance (p= 0.002 and p= 0.03, respectively) for Candida spp. strains isolated from IUD string samples. The overall resistance rates of C.albicans and non-albicans Candida spp. against fluconazole, were determined as 15% and 54.2%, respectively, while those rates were 24.2% and 68.7%, respectively, against itraconazole. MIC value of amphotericin B for all of the Candida spp. isolates was ≤ 1.5 µg/ml. In conclusion, the data obtained from this study revealed that Candida spp. May lead to vaginal infections by inducing biofilm formation in IUD strings and these biofilms may be related to resistance to antifungal agents. Thus, women using IUDs should be followed-up periodically for the development of biofilms in their IUD strings.

摘要

宫内节育器(IUD)应用是一种广泛使用的有效、安全且经济的计划生育方法。然而,使用IUD可能会导致阴道生态系统发生某些变化,并可能扰乱微生物群落,导致各种机会性病原体微生物的定植增加。本研究的目的是:(i)检测从使用IUD的女性阴道和IUD尾丝样本中分离出的念珠菌属的生物膜产生特征,以及(ii)研究生物膜产生与抗真菌耐药性之间的关系。共有250名因阴道症状(分泌物和瘙痒)入住妇科门诊的女性纳入本研究(平均年龄:34.4±7.6岁)。患者植入CuT380a型IUD的平均时间为59.8±42.4个月。在不取出IUD的情况下,通过剪断获取尾丝样本,同时也采集阴道拭子样本。分离出的念珠菌属通过常规方法和API 20C AUX(法国生物梅里埃公司)系统进行鉴定。根据CLSI指南,采用肉汤微量稀释法测定氟康唑、伊曲康唑和两性霉素B的最低抑菌浓度(MIC)。通过结晶紫染色和XTT还原试验评估生物膜形成,在两种方法中均产生阳性结果的分离株被视为生物膜产生菌。在本研究中,念珠菌属分别从33.2%(83/250)的阴道样本和34%(85/250)的IUD尾丝样本中分离得到,白色念珠菌是最常检测到的菌种(样本中分别为54株和66株)。阴道分离株的体外生物膜总形成率为25%(21/83),IUD尾丝分离株为44.7%(38/85)。阴道白色念珠菌分离株的生物膜形成率显著低于阴道非白色念珠菌属(分别为14.8%和

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