Coban Ahmet Yilmaz, Ciftci Alper, Onuk Ertan Emek, Erturan Zayre, Tanriverdi Cayci Yeliz, Durupinar Belma
Ondokuz Mayls Universitesi Tip Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dali, Samsun.
Mikrobiyol Bul. 2009 Oct;43(4):563-73.
Pseudomonas aeruginosa is a frequent cause of respiratory infections in cystic fibrosis (CF) patients. P. aeruginosa strains isolated from these patients have often a mucoid phenotype at advanced disease. This mucoid structure contains a dense amount of alginate type polysaccharide which facilitates bacterial attachment to lung epithelia and provides protection from the immune system due to biofilm formation. The aims of this study were to investigate the biofilm formation and the relation of this property with genotype and antibiotic susceptibilities of P. aeruginosa strains isolated from CF patients. The biofilm formation was determined by using the Congo Red agar and Christensen methods. RAPD-PCR (Random amplification of polymorphic DNA polymerase chain reaction) and disc diffusion methods were used for genotyping and antibiotic susceptibility testing, respectively. Biofilm production was found positive in 33.3% (20/60) of P. aeruginosa tested. While 9 of these 20 isolates were of mucoid colony morphotype, among the 40 biofilm negative isolates mucoid colony was detected in 16 of them. RAPD genotyping based on 70% similarity yielded 19 (A-S) clusters and subtypes related to five of these clusters (K1, K2, N1, N2, Q1, Q2, R1, R2, S1, S2) making up a total of 24 genotypes. Nine of these genotypes composed of biofilm positive isolates and 15 were biofilm negative ones. Most of the biofilm positive strains belonged to K1 (n = 5) and K2 (n = 6) genotypes while biofilm negative isolates were in the L (n = 8) and O (n = 7) genotypes. The comparison of antibiotic susceptibilities in both groups revealed no statistically significant difference (p > 0.0%). However, highest rate of resistance was detected for tobramycin and lowest rate for piperacillin/tazobactam. The data obtained from this study indicated that biofilm negative and positive P. aeruginosa isolates clustered in different groups. These results should be supported with larger scale multi-center studies which may provide information about P. aeruginosa dynamics in CF lungs.
铜绿假单胞菌是囊性纤维化(CF)患者呼吸道感染的常见病因。从这些患者中分离出的铜绿假单胞菌菌株在疾病晚期通常具有黏液样表型。这种黏液样结构含有大量藻酸盐型多糖,有助于细菌附着于肺上皮细胞,并由于生物膜形成而提供对免疫系统的保护。本研究的目的是调查从CF患者中分离出的铜绿假单胞菌菌株的生物膜形成情况,以及该特性与基因型和抗生素敏感性之间的关系。通过使用刚果红琼脂和克里斯滕森方法测定生物膜形成。分别使用RAPD-PCR(随机扩增多态性DNA聚合酶链反应)和纸片扩散法进行基因分型和抗生素敏感性测试。在测试的铜绿假单胞菌中,33.3%(20/60)的菌株生物膜产生呈阳性。这20株分离株中有9株为黏液样菌落形态型,在40株生物膜阴性分离株中,有16株检测到黏液样菌落。基于70%相似性的RAPD基因分型产生了19个(A-S)簇以及与其中五个簇(K1、K2、N1、N2、Q1、Q2、R1、R2、S1、S2)相关的亚型,总共构成24种基因型。这些基因型中有9种由生物膜阳性分离株组成,15种为生物膜阴性分离株。大多数生物膜阳性菌株属于K1(n = 5)和K2(n = 6)基因型,而生物膜阴性分离株属于L(n = 8)和O(n = 7)基因型。两组抗生素敏感性的比较未显示出统计学上的显著差异(p > 0.0%)。然而,检测到对妥布霉素的耐药率最高,对哌拉西林/他唑巴坦的耐药率最低。本研究获得的数据表明,生物膜阴性和阳性的铜绿假单胞菌分离株聚集在不同的组中。这些结果应得到更大规模多中心研究的支持,这些研究可能提供有关CF肺部铜绿假单胞菌动态的信息。