Manaka Akihiro, Tokue Yutaka, Murakami Masami
Department of Clinical Laboratory Medicine, Gunma University Graduate School of Medicine, Maebashi, Gunma Japan.
Infection Control and Prevention Center, Gunma University Hospital, 3-39-15 Showa-machi, Maebashi, Gunma Japan.
J Pharm Health Care Sci. 2017 Jan 19;3:8. doi: 10.1186/s40780-017-0074-y. eCollection 2017.
Nosocomial infection is one of the most common complications within health care facilities. Certain studies have reported outbreaks resulting from contaminated hospital environments. Although the identification of bacteria in the environment can readily be achieved using culturing methods, these methods detect live bacteria. Sequencing of the 16S ribosomal RNA (16S rRNA) gene is recognized to be effective for bacterial identification. In this study, we surveyed wards where drug-resistant bacteria had been isolated and compared conventional culture methods with 16S rRNA gene sequencing methods.
Samples were collected using sterile swabs from two wards (northern and southern) at Gunma University Hospital contaminated by sp.. We extracted DNA directly from the swabs. Following extraction, the DNA was amplified using polymerase chain reaction (PCR). The PCR products were cloned using the plasmid vector. The plasmid DNA were sequenced, and identification were performed using database. 16S rRNA gene sequence analyses were compared conventional culture methods.
In the northern ward, sp. was detected from only two of 14 samples using the culture method. In contrast, 16S rRNA gene sequencing analysis detected sp. from seven of 14 samples. Drug-resistant sp. was isolated from bathrooms of the southern ward and was detected from four of seven samples using the culture method in comparison with six of seven samples by 16S rRNA gene sequencing analysis.
Molecular biological analysis showed a higher sensitivity to detect specific bacteria and detected a greater number of species than the culture method. Our results suggest that 16S rRNA gene sequencing analysis is useful to identify range of contamination which were not found in conventional culture method. When a nosocomial outbreak cannot be adequately controlled, molecular biological analysis may serve as a useful tool for environmental surveys in hospitals.
医院感染是医疗机构中最常见的并发症之一。某些研究报告了由医院环境污染导致的感染暴发。尽管使用培养方法可以很容易地鉴定环境中的细菌，但这些方法只能检测活细菌。16S核糖体RNA（16S rRNA）基因测序被认为对细菌鉴定有效。在本研究中，我们对分离出耐药菌的病房进行了调查，并将传统培养方法与16S rRNA基因测序方法进行了比较。
使用无菌拭子从群马大学医院的两个病房（北部和南部）采集样本，这些病房被……污染。我们直接从拭子中提取DNA。提取后，使用聚合酶链反应（PCR）扩增DNA。使用质粒载体克隆PCR产物。对质粒DNA进行测序，并使用数据库进行鉴定。将16S rRNA基因序列分析与传统培养方法进行比较。
在北部病房，使用培养方法仅从14个样本中的2个检测到……。相比之下，16S rRNA基因测序分析从14个样本中的7个检测到……。从南部病房的浴室中分离出耐药……，使用培养方法从7个样本中的4个检测到，而通过16S rRNA基因测序分析从7个样本中的6个检测到。
分子生物学分析显示出比培养方法更高的检测特定细菌的灵敏度，并检测到更多种类的细菌。我们的结果表明，16S rRNA基因测序分析有助于识别传统培养方法未发现的污染范围。当医院感染暴发无法得到充分控制时，分子生物学分析可能是医院环境调查的有用工具。
