Preparation of quality control materials for SARS-CoV-2 variants based on MS2 phage virus-like particles
10.13602/j.cnki.jcls.2025.10.11
- VernacularTitle:基于MS2噬菌体病毒样颗粒的新型冠状病毒变异株质控品的制备
- Author:
Ran ZHAO
1
;
Yingwei CHEN
;
Chengxiang CHU
;
Zhongqiang HUANG
;
Weijie DING
;
Xueliang WANG
Author Information
1. 上海市临床检验中心,上海 200126;上海市实验医学研究院,上海 200126
- Publication Type:Journal Article
- Keywords:
SARS-CoV-2 variant;
MS2 phage;
virus-like particle;
quality control material;
protein purification
- From:
Chinese Journal of Clinical Laboratory Science
2025;43(10):773-779
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a variety of quality control(QC)materials for SARS-CoV-2 variants as an addition to the conven-tional SARS-CoV-2 nucleic acid QC products for the laboratory detection of mutant strains by optimizing the preparation and purification process of MS2 phage virus-like particle(VLP)technique,and evaluate their performances.Methods The typical mutation sequence fragments or full length S genes were designed and synthesized according to the genomic information of SARS-CoV-2 variants.Then,they were inserted into the downstream of maturase gene,coat protein and the pac-site of MS2 phage to construct a series of recombi-nant expression vectors.After induced by the prokaryotic expression system,the VLP products were purified through the polyethylenei-mine precipitation,ultrafiltration,nuclease digestion,and gel filtration chromatography.The obtained VLP were validated by the nucle-ic acid electrophoresis,protein electrophoresis,protein concentration determination,and fluorescence PCR,and their performances such as nucleic acid residue and stability were also evaluated.Results A total of 10 kinds of VLP containing the targeted sequences of the gene to be tested were prepared.The length of the foreign sequence wrapped in them ranged from 297 bp to 3 822 bp,which could be combined into a variety of QC materials for the mutation detection of different SARS-CoV-2 variants.The prepared VLP QC materials could not be effectively amplified without nucleic acid extraction or reverse transcription steps during the routine nucleic acid detection.The simulated QC samples remained stable after repeated freeze-thaw cycles.They could be stored stably for 2 months at 25 ℃ and 4 weeks at 37 ℃.Conclusion The established preparation and combined purification process of VLP QC materials can encapsulate vari-ous exogenous nucleic acid sequences with different lengths into the viral coat protein to form VLP,with high production efficiency.The VLP QC products prepared by the above process have stable performance and almost no residual exogenous nucleic acid,which can ef-fectively meet clinical requirements and ensure the quality of laboratory testing.
