Department of Microbiology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Eye Contact Lens. 2009 Mar;35(2):81-7. doi: 10.1097/ICL.0b013e318199b044.
Disposable soft contact lenses that are commonly used after laser refractive surgery are known to be colonized by bacteria and play a key role in Bacterial Keraitis (BK) pathogenesis. Coagulase-negative staphylococci (CoNS) have been found to be the most common pathogen involved in this postoperative infection. In this study a rapid and a simple assay was developed for studying attachment and accumulation of CoNS on soft contact lenses in vitro using [3H] thymidine.
Thirty-five isolates of CoNS were obtained from 27 laser refractive surgery patients. Twenty-five of these thirty-five CoNS were isolated in multiple cultures. Ten CoNS were isolated in cultures from patients who underwent reoperation. The assay was optimized using a biofilm-producing strain, S. epidermidis RP62A, which was subcultured overnight at 37 degrees C on blood agar medium. Quantitative determination of biofilm production was tested. Presence of the genes icaADB and icaD was determined in all isolates. All isolates were biochemically analyzed using the Phene Plate (PhP) system modified for typing of CoNS. The CoNS isolates were further characterized to species level using ID32Staph.Mann-Whitney rank sum test and chi-square test were used to identify statistical differences in adherence, index, antibiotic susceptibility patterns, and biofilm production or presence of the ica operon between clinically significant isolates and non-postoperative BK isolates.
No differences in attachment and accumulation were found between isolates causing BK after laser refractive surgery and contaminant isolates. In addition, there were no differences in the distribution of the ica operon between the two groups, as determined by polymerase chain reaction. Nevertheless, the ability to produce biofilm was found to be present significantly more frequently among BK isolates than among non-postoperative BK isolates.
This study shows that the method using radioactive thymidine to analyze adherence of CoNS to soft contact lenses enables detection of differences in the adherence patterns of individual isolates.
激光屈光手术后常用的一次性软性隐形眼镜已被证实会被细菌定植,并在细菌性角膜炎(BK)发病机制中起关键作用。凝固酶阴性葡萄球菌(CoNS)已被发现是与这种术后感染相关的最常见病原体。在这项研究中,我们开发了一种快速而简单的方法,使用[3H]胸苷研究 CoNS 在体外对软性隐形眼镜的附着和积累。
从 27 名激光屈光手术患者中获得了 35 株 CoNS 分离株。这 35 株 CoNS 中有 25 株是从多次培养中分离出来的。10 株 CoNS 是从接受再次手术的患者的培养物中分离出来的。该方法使用生物膜产生菌株 S. epidermidis RP62A 进行了优化,该菌株在血琼脂培养基上于 37°C 过夜培养。测试了生物膜产生的定量测定。在所有分离株中均检测到基因 icaADB 和 icaD 的存在。所有分离株均使用改良的 Phene Plate(PhP)系统进行了生化分析,该系统用于 CoNS 的分型。使用 ID32Staph.Mann-Whitney 秩和检验和卡方检验进一步将 CoNS 分离株鉴定为种水平。
在导致激光屈光手术后 BK 的分离株和污染分离株之间,在附着和积累方面未发现差异。此外,通过聚合酶链反应,在这两组之间,ica 操纵子的分布也没有差异。然而,与非术后 BK 分离株相比,生物膜产生能力在 BK 分离株中更为常见。
本研究表明,使用放射性胸腺嘧啶分析 CoNS 对软性隐形眼镜的附着的方法可以检测到各个分离株附着模式的差异。