Establishing integrated system-based point-of-care testing method for genotyping of respiratory syncytial virus
10.3760/cma.j.cn112309-20240530-00183
- VernacularTitle:基于一体化系统的呼吸道合胞病毒分型即时检验检测技术的建立及应用
- Author:
Fei DENG
1
;
Liguo ZHU
;
Ke XU
;
Xian QI
;
Huiyan YU
;
Shenjiao WANG
;
Changjun BAO
Author Information
1. 江苏省疾病预防控制中心急性传染病防制所，南京 210009
- Keywords:
Respiratory syncytial virus;
Genotyping;
POCT reagent;
Integration
- From:
Chinese Journal of Microbiology and Immunology
2024;44(8):660-664
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To develop an integrated point-of-care testing (POCT) reagent for genotying respiratory syncytial virus (RSV) and evaluate its performance.Methods:Specific primers and probes were designed based on the conserved sequences of the genomes of RSV A and B as well as ON1 and BA9 genotypes. The PCR reaction system and conditions were optimized. The vitrification technology of reagents and multiplex detection platform were integrated to develop the RSV genotyping POCT reagent. The sensitivity, specificity, reproducibility, and clinical performance of the product were then evaluated.Results:The sensitivity of the developed integrated RSV genotyping POCT reagent reached 500 copies/ml. It exhibited good specificity with no cross-reaction with clinically similar pathogens. The coefficient of variation of Ct values for both inter-batch and intra-batch reproducibility was less than 5%, indicating good reproducibility. In testing 53 clinical samples, the detection results showed high consistency and concordance with the reference reagent, with a positive concordance rate of up to 98.11%.Conclusions:The developed integrated RSV genotyping POCT reagent incorporates nucleic acid extraction, purification, and detection into a single process, achieving a "sample in, result out" workflow. It is simple to operate and provides accurate, reliable, and stable detection results. This product can be used for the genotyping of RSV A and B in POCT, offering support for the prevention, control, and diagnosis of RSV.
