摘要
目的 采用自建微流控-芯片法检测系统对本实验室于2021年5月至2022年8月期间收集保存的布鲁氏菌阳性菌株、阳性血清、阴性血清进行检测,验证自建系统检测布鲁氏菌核酸的主要性能是否满足相关标准.方法 依据CNAS-GL039:2019《分子诊断检验程序性能验证指南》、《实验室自建分子诊断项目基本要求专家共识》、《布鲁菌病诊疗专家共识》相关要求,对自建微流控-芯片法检测系统的符合率、重复性、检出限、交叉反应、抗干扰能力、敏感性及特异性进行验证及评价.结果 自建检测系统与经血培养鉴定为布鲁氏菌阳性菌株、体检者阴性标本检测结果的阳性符合率为100％,阴性符合率为100％,总符合率为100％;1.50×108CFU/mL、500 CFU/mL布鲁氏菌阳性标本重复性检出率均为100％;检出限浓度(1000 copies/mL)企业参考品20次重复检测结果均为阳性;七种交叉反应病原体标本检测中,布鲁氏菌核酸结果均为阴性;在混有干扰物质的弱阳性布鲁氏菌标本检测中,布鲁氏菌核酸结果均为阳性;在布鲁氏菌感染检测中的敏感性和特异性分别为89.88％、95.74％.结论 微流控-芯片法检测系统的符合率、重复性、检出限、抗干扰能力、交叉反应、敏感性及特异性性能指标均符合相关标准及检测要求,可用于临床检测工作.
Abstract
Objective To detect positive strains,and positive/negative serum samples of Brucella collected and stored in our laboratory from May 2021 to August 2022,to verify whether the self-built system met the standards for detecting Brucella nucleic acid. Methods The selt-built microfluidic chip detection system was evaluated and verified for its coincidence rate,repeatability,detection-limit,cross-reaction,anti-interference ability,sensitivity,and specificity. This evaluation was conducted according to the CNAS-GL039:2019, Expert Consensus on Basic Requirements of Laboratory Self-Built Molecular Diagnostic Projects and Expert Consensus on Brucellosis Diagnosis and Treatment. Results The selt-built microfluidic chip detection system showed a 100％coincidence rate for detecting positive Brucella strains,and negative samples from physical examiners when compare to blood culture. The repeatability of detecting positive Brucella samples with 1.50 × 108 CFU/mL and 500 CFU/mL was 100％. The detection-limit concentration(1000 copies/mL)of the enterprise reference material was successfully detected in all 20 repeated tests. No Brucella nucleic acids were detected in the detection of seven cross-reaction pathogens. However,Brucella nucleic acids were detected in the detection of weakly positive Brucella samples mixed with interfering substances. The sensitivity and specificity of detecting Brucella were 89.88％and 95.74％,respectively. Conclusion The selt-built microfluidic chip detection system demonstrated a high coincidence rate,repeatability,detection-limit,and anti-interference ability,lack of cross-reaction,and satisfactory sensitivity and specificity. Therefore,this system can be applied for clinical detection.
基金项目
内蒙古自治区卫生健康科技计划(202201609)
维普
万方数据