Characterization and complete genome sequence of highly lytic phage active against methicillin-resistant Staphylococcus aureus (MRSA) isolated from Egypt.

IF 4 3区 医学 Q2 VIROLOGY
Abeer K Abd El-Tawab, B A Othman, A Sharaf, Samar S El-Masry, T F El-Arabi
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引用次数: 0

Abstract

Background: Methicillin-Resistant Staphylococcus aureus (MRSA) is one of the most resistant bacteria to antibiotics. S. aureus is an important, widespread pathogen that can cause a variety of infectious diseases in humans and animals. Phages have been recognized as natural, safe, highly specific and effective alternatives agents to antibiotics for preventing and treating bacterial infections caused by MRSA. Therefore, this study aims at the characterization of a novel isolated lytic phage, vB_SauP_ASUmrsa123.

Methods: Isolates of Staphylococcus aureus MRSA were obtained on Mannitol Salt Agar and Baird Parker Agar plates and confirmed using VITEK 2. Sewage and clinical samples were used to isolate specific phages for S. aureus MRSA, and plaque assays were used for host range determination on Luria-Bertani (LB) media. The phage morphology of the isolated phage was determined by transmission electron microscopy. The phage's whole genome sequencing was identified.

Results: A total of 25 isolates of Staphylococci were obtained from different clinical sources and showed typical colonies on Baird-Parker and Mannitol Salt Agar plates. The VITEK 2 automated system revealed that all 25 isolates were confirmed as S. aureus (MRSA). Two of the most antibiotics-resistant isolates were further confirmed using 16S ribosomal RNA sequencing. A lytic phage was detected against the MRSA isolates tested In Vitro, namely vB_SauP_ASUmrsa123. The phage belonged to Rountreeviridae family based on morphological properties observed by TEM and the host range of the isolated phage was tested on the 25 clinical MRSA isolates in Vitro. The one-step growth curve of the isolated phage showed that the latent period was about 55 min, and the burst size was estimated at 167 PFU. The whole genome sequencing and annotation of genes revealed that phage vB_SauP_ASUmrsa123 contained a linear dsDNA with a size of about 17,155 bp with predicted 24 ORFs. Analysis of its genome provides valuable information approximately the variety of phages belonging to the staphylococcal phages class I.

Conclusion: A lytic Podo Phage vB_SauP_ASUmrsa123 was identified against S. aureus MRSA isolates and its genome was sequenced. The phage was found to be eligible for potential application in biocontrol.

对埃及分离出的耐甲氧西林金黄色葡萄球菌(MRSA)具有活性的高溶解性噬菌体的特征和完整基因组序列。
背景:耐甲氧西林金黄色葡萄球菌(MRSA耐甲氧西林金黄色葡萄球菌(MRSA)是对抗生素耐药性最强的细菌之一。金黄色葡萄球菌是一种重要的、广泛传播的病原体,可导致人类和动物的各种传染性疾病。噬菌体被认为是天然、安全、高度特异性和有效的抗生素替代药剂,可用于预防和治疗由 MRSA 引起的细菌感染。因此,本研究旨在鉴定一种新分离的溶菌噬菌体 vB_SauP_ASUmrsa123:方法:在甘露醇盐琼脂和贝尔德-帕克琼脂平板上获得金黄色葡萄球菌 MRSA 的分离物,并用 VITEK 2 进行确认。通过透射电子显微镜确定了分离出的噬菌体的形态。对噬菌体进行了全基因组测序:结果:从不同临床来源共分离到 25 株葡萄球菌,在 Baird-Parker 和甘露醇盐琼脂平板上显示出典型菌落。VITEK 2 自动系统显示,所有 25 个分离物均被确认为金黄色葡萄球菌(MRSA)。其中两个抗生素耐药性最强的分离株通过 16S 核糖体 RNA 测序得到了进一步确认。针对体外检测的 MRSA 分离物检测到了一种溶菌噬菌体,即 vB_SauP_ASUmrsa123。根据 TEM 观察到的形态特征,该噬菌体属于 Rountreeviridae 科。分离噬菌体的一步生长曲线显示,潜伏期约为 55 分钟,迸发量估计为 167 PFU。全基因组测序和基因注释显示,vB_SauP_ASUmrsa123噬菌体含有线性dsDNA,大小约为17,155 bp,预测有24个ORFs。对其基因组的分析为了解属于葡萄球菌噬菌体 I 类的噬菌体种类提供了有价值的信息:结论:针对金黄色葡萄球菌 MRSA 分离物鉴定出了一种溶菌性 Podo 噬菌体 vB_SauP_ASUmrsa123,并对其基因组进行了测序。发现该噬菌体有可能应用于生物控制。
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来源期刊
Virology Journal
Virology Journal 医学-病毒学
CiteScore
7.40
自引率
2.10%
发文量
186
审稿时长
1 months
期刊介绍: Virology Journal is an open access, peer reviewed journal that considers articles on all aspects of virology, including research on the viruses of animals, plants and microbes. The journal welcomes basic research as well as pre-clinical and clinical studies of novel diagnostic tools, vaccines and anti-viral therapies. The Editorial policy of Virology Journal is to publish all research which is assessed by peer reviewers to be a coherent and sound addition to the scientific literature, and puts less emphasis on interest levels or perceived impact.
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