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利用抗原组技术从重症监护病房工作人员和感染患者血清中鉴定医院获得性鲍曼不动杆菌临床分离株的抗原

Identification of antigens from nosocomial Acinetobacter baumannii clinical isolates in sera from ICU staff and infected patients using the antigenome technique.

作者信息

Nafarieh Tina, Bandehpour Mojgan, Hashemi Ali, Taheri Sodabeh, Yardel Vahid, Jamaati Hamidreza, Moosavi Seyed Mahdi, Mosaffa Nariman

机构信息

Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

World J Microbiol Biotechnol. 2017 Sep 30;33(10):189. doi: 10.1007/s11274-017-2355-x.

Abstract

Nosocomial infections with a bacterial origin are considered one of the most dangerous threats to global health. Among the causes of these infections, Acinetobacter baumannii is playing a significant role, and the present study aimed‏ to determine the immunogenic proteins of this bacteria. Clinical isolates of A. baumannii were obtained from positive sputum cultures of intensive care unit (ICU) patients confirmed by Polymerase chain reaction (PCR) of the OXA-51 gene, and sera was obtained from 20 colonized patients. In addition, 20 and 30 serum samples were collected from ICU nurses and healthy controls, respectively. All the samples were screened in the presence of antibodies against A. baumannii by enzyme-linked immunosorbent assay (ELISA). IgG purified from the serum samples by affinity chromatography was used to isolate the bacteria by the Magnetic-activated cell sorting (MACS) procedure. After the bacteria were cultured, the identified antigen proteins were studied by western blotting and Mass spectrometry (MS). The MS results were analyzed with MASCOT software and revealed a 35 KD protein, which corresponds to outer membrane protein A (OmpA) of A. baumannii, a 25 KD band, which is a carbapenem-associated resistance protein precursor, and a 60 KD protein band, identified as a stress-induced bacterial acidophilic repeat motif protein. According to the properties of immunogen antigens and bio informatics tools, the outer membrane proteins (OMPs) can be used as a vaccine candidate in animal models.

摘要

源自细菌的医院感染被认为是对全球健康最危险的威胁之一。在这些感染的病因中,鲍曼不动杆菌起着重要作用,本研究旨在确定该细菌的免疫原性蛋白。鲍曼不动杆菌的临床分离株取自重症监护病房(ICU)患者经OXA - 51基因聚合酶链反应(PCR)确认的阳性痰培养物,并从20名定植患者中采集血清。此外,分别从ICU护士和健康对照中收集了20份和30份血清样本。所有样本均通过酶联免疫吸附测定(ELISA)在抗鲍曼不动杆菌抗体存在的情况下进行筛选。通过亲和色谱从血清样本中纯化的IgG用于通过磁激活细胞分选(MACS)程序分离细菌。细菌培养后,通过蛋白质印迹和质谱(MS)研究鉴定出的抗原蛋白。用MASCOT软件分析MS结果,发现一条35 KD的蛋白条带,对应于鲍曼不动杆菌的外膜蛋白A(OmpA),一条25 KD的条带,是一种碳青霉烯类耐药蛋白前体,还有一条60 KD的蛋白条带,鉴定为应激诱导的细菌嗜酸重复基序蛋白。根据免疫原抗原的特性和生物信息学工具,外膜蛋白(OMPs)可作为动物模型中的疫苗候选物。

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