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  

Establishing integrated system-based point-of-care testing method for genotyping of respiratory syncytial virus

作　　者：邓斐[1] 朱立国[1] 许可[1] 祁贤[1] 余慧燕[1] 王慎骄[1] 鲍倡俊[1] Deng Fei;Zhu Liguo;Xu Ke;Qi Xian;Yu Huiyan;Wang Shenjiao;Bao Changjun(Acute Infectious Disease Prevention and Control Institute,Jiangsu Provincial Center for Disease Control and Prevention,Nanjing 210009,China)

机构地区：[1]江苏省疾病预防控制中心急性传染病防制所,南京210009

出　　处：《中华微生物学和免疫学杂志》2024年第8期660-664,共5页Chinese Journal of Microbiology and Immunology

基　　金：江苏省社会发展项目(BE2021739);江苏省第六期"333工程"重点行业领域人才项目。

摘　　要：目的研发一款一体化呼吸道合胞病毒(RSV)分型即时检验(point-of-care testing,POCT)检测试剂并评估其性能。方法以RSV A和B亚型及ON1和BA9基因型的基因组保守序列设计特异性的引物和探针,优化PCR反应体系和条件,整合试剂玻璃化技术和多重检测平台,开发RSV分型POCT检测试剂,并对该产品的敏感度、特异度、重复性和临床性能进行评估。结果一体化RSV分型POCT检测试剂敏感度可达500拷贝/ml,特异度好,与临床上表现相似的病原体无交叉反应,试剂批间和批内重复性Ct值的变异系数均<5%,具有较好的重复性,检测试剂应用于53例临床样本测试,检测结果与对比试剂具有较高的一致性和符合率,阳性符合率高达98.11%。结论本研究开发的一体化RSV分型POCT检测试剂集成了"核酸提取-纯化-检测",实现了"样本进,结果出",操作简单,检测结果准确、可靠、稳定,可用于RSV A和B亚型的分型即时检验,为RSV的防控和诊疗提供助力。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.

关 键 词：呼吸道合胞病毒 分型 POCT检测试剂 一体化 

分 类 号：R511[医药卫生—内科学] R446.5[医药卫生—临床医学]