Rahimzadeh Golnar, Gill Pooria, Rezai Mohammad Sadegh
Pediatric Infectious Diseases Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Nanomedicine Group, Immunogenetics Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Avicenna J Med Biotechnol. 2020 Apr-Jun;12(2):85-90.
Emergence and prevalence of multi drug resistance strains such as Methicillin-Resistant (MRSA) call for new antibacterial option. Endolysins as a new option is suggested. The phage display technique is suggested for production of recombinant endolysins. The recombinant endolysins displayed nano phages specifically lysis bacteria, which penetrate to the depth of tissue and the effective dose is reduced.
gene was ligated in T7Select vector arms in T7Select10-3b cloning kit. To produce recombinant nano phages, ligation reaction was added directly to the packaging extract. Recombinant nano phages were amplified by Double Layer Agar assay (DLA). The recombinant nano phages were characterized using TEM. Size of recombinant nano phages was determined using DLS. The spot test was performed to confirm CHAPk -displayed on the surface of nano phages. The turbidimetry was used to investigate lytic activity of recombinant nano phages against MRSA ATCC No. 33591.
The results showed recombinant nano phages belonged to order Caudovirales and family Podoviridae with titer 2×10 . According to the results of DLS, size of recombinant nano phages was 71 nm. Formation inhibition zone confirmed the presence of CHAPk on the surface of nano phage phenotypically. The turbidimetry showed lytic activity recombinant nano phages against MRSA after 5 .
This study suggests that CHAPk -displayed nano phages can be effective in MRSA infections.
耐甲氧西林金黄色葡萄球菌（MRSA）等多重耐药菌株的出现和流行需要新的抗菌选择。内溶素作为一种新选择被提出。噬菌体展示技术被建议用于生产重组内溶素。展示重组内溶素的纳米噬菌体可特异性裂解细菌，其可渗透到组织深处且有效剂量降低。
将基因连接到T7Select10 - 3b克隆试剂盒中的T7Select载体臂上。为生产重组纳米噬菌体，将连接反应直接加入包装提取物中。通过双层琼脂试验（DLA）扩增重组纳米噬菌体。使用透射电子显微镜（TEM）对重组纳米噬菌体进行表征。使用动态光散射（DLS）测定重组纳米噬菌体的大小。进行斑点试验以确认纳米噬菌体表面展示的CHAPk。使用比浊法研究重组纳米噬菌体对金黄色葡萄球菌ATCC编号33591的裂解活性。
结果显示重组纳米噬菌体属于有尾噬菌体目和短尾噬菌体科，滴度为2×10 。根据DLS结果，重组纳米噬菌体的大小为71纳米。形成抑菌圈从表型上证实了纳米噬菌体表面存在CHAPk。比浊法显示重组纳米噬菌体在5 后对MRSA具有裂解活性。
本研究表明展示CHAPk的纳米噬菌体对MRSA感染可能有效。
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