Seng Rathanin, Kitti Thawatchai, Thummeepak Rapee, Kongthai Phattaraporn, Leungtongkam Udomluk, Wannalerdsakun Surat, Sitthisak Sutthirat
Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand.
Faculty of Oriental Medicine, Chiang Rai College, Chiang Rai, Thailand.
PLoS One. 2017 Aug 31;12(8):e0184172. doi: 10.1371/journal.pone.0184172. eCollection 2017.
Methicillin-resistant coagulase negative staphylococci (MR-CoNS) are the major cause of infectious diseases because of their potential ability to form biofilm and colonize the community or hospital environments. This study was designed to investigate the biofilm producing ability, and the presence of mecA, icaAD, bap and fnbA genes in MR-CoNS isolates. The MR-CoNS used in this study were isolated from various samples of community environment and five wards of hospital environments, using mannitol salt agar (MSA) supplemented with 4 μg/ml of oxacillin. The specie level of Staphylococcus haemolyticus, Staphylococcus epidermidis, Staphylococcus hominis and Staphylococcus warneri was identified by specific primers of groESL (S. haemolyticus), rdr (S. epidermidis) and nuc (S. hominis and S. warneri). The remainder isolates were identified by tuf gene sequencing. Biofilm production was determined using Congo red agar (CRA) and Microtiter plate (MTP) assay. The mecA and biofilm associated genes (icaAD, fnbA and bap) were detected using PCR method. From the 558 samples from community and hospital environments, 292 MR-CoNS were isolated (41 from community environments, and 251 from hospital environments). S. haemolyticus (41.1%) and S. epidermidis (30.1%) were the predominant species in this study. Biofilm production was detected in 265 (90.7%) isolates by CRA, and 260 (88.6%) isolates were detected by MTP assay. The staphylococci isolates derived from hospital environments were more associated with biofilm production than the community-derived isolates. Overall, the icaAD and bap genes were detected in 74 (29.5%) and 14 (5.6%) of all isolates from hospital environments. When tested by MTP, the icaAD gene from hospital environment isolates was associated with biofilm biomass. No association was found between bap gene and biofilm formation. The MR-CoNS isolates obtained from community environments did not harbor the icaAD and bap genes. Conversely, fnbA gene presented in MR-CoNS isolated from both community and hospital environments. The high prevalence of biofilm producing MR-CoNS strains demonstrated in this study indicates the persisting ability in environments, and is useful in developing prevention strategies countering the spread of MR-CoNS.
耐甲氧西林凝固酶阴性葡萄球菌(MR-CoNS)是传染病的主要病因,因为它们具有形成生物膜并在社区或医院环境中定殖的潜在能力。本研究旨在调查MR-CoNS分离株的生物膜形成能力,以及mecA、icaAD、bap和fnbA基因的存在情况。本研究中使用的MR-CoNS是从社区环境的各种样本以及医院环境的五个病房中分离出来的,使用添加了4μg/ml苯唑西林的甘露醇盐琼脂(MSA)。通过groESL(溶血葡萄球菌)、rdr(表皮葡萄球菌)和nuc(人葡萄球菌和沃氏葡萄球菌)的特异性引物鉴定溶血葡萄球菌、表皮葡萄球菌、人葡萄球菌和沃氏葡萄球菌的种属水平。其余分离株通过tuf基因测序进行鉴定。使用刚果红琼脂(CRA)和微量滴定板(MTP)试验测定生物膜的产生。使用PCR方法检测mecA和生物膜相关基因(icaAD、fnbA和bap)。从社区和医院环境的558个样本中,分离出292株MR-CoNS(41株来自社区环境,251株来自医院环境)。溶血葡萄球菌(41.1%)和表皮葡萄球菌(30.1%)是本研究中的主要菌种。通过CRA检测到265株(90.7%)分离株产生生物膜,通过MTP试验检测到260株(88.6%)分离株产生生物膜。源自医院环境的葡萄球菌分离株比源自社区的分离株与生物膜产生的相关性更强。总体而言,在所有源自医院环境的分离株中,74株(29.5%)检测到icaAD基因,14株(5.6%)检测到bap基因。通过MTP检测时,源自医院环境分离株的icaAD基因与生物膜生物量相关。未发现bap基因与生物膜形成之间存在关联。从社区环境中获得的MR-CoNS分离株未携带icaAD和bap基因。相反,fnbA基因存在于从社区和医院环境中分离出的MR-CoNS中。本研究中显示的产生生物膜的MR-CoNS菌株的高流行率表明其在环境中的持续存在能力,有助于制定应对MR-CoNS传播的预防策略。