摘要
目的探讨重组酶聚合酶扩增(RPA)技术用于脑脊液(CSF)中肺炎链球菌检测的价值。方法利用RPA技术检测CSF中肺炎链球菌。根据肺炎链球菌LepA基因设计RPA反应引物,收集临床确诊和疑似的52例细菌性脑膜炎患者为研究对象,采集CSF标本并提取DNA,采用RPA技术对临床CSF进行LepA基因扩增检测。结果52例患者的CSF标本经微生物培养有19例(36.5%)为阳性,阳性标本中有7例检出肺炎链球菌,这7例患者CSF标本的RPA检测结果均为阳性。微生物培养为阴性的33例(63.5%)患者中有7例患者CSF标本的RPA检测结果为阳性。共14例患者CSF标本的RPA检测结果为阳性,从中选取10例患者CSF标本的RPA扩增产物经纯化后,与肺炎链球菌LepA基因目的片段进行核酸序列比对,其比对结果一致。结论RPA技术可用于肺炎链球菌性脑膜炎病原菌的快速检测。
Objective To investigate the value of recombinase polymerase amplification(RPA)assay in detecting of Streptococcus pneumoniae in cerebrospinal fluid(CSF).Methods RPA was used to detect Streptococcus pneumoniae in CSF.According to the LepA gene of Streptococcus pneumoniae,RPA reaction primers were designed,52 cases of clinically confirmed and suspected bacterial meningitis were collected as research objects,CSF samples were collected and DNA was extracted,and LepA gene amplification of clinical CSF was detected by RPA technology.Results Among the 52 patients,19(36.5%)of the CSF samples were positive by microbial culture,and 7 of the positive samples were positive for Streptococcus pneumoniae.The RPA test results of the CSF samples of these 7 patients were positive.The CSF samples of 33 patients(63.5%)were negative by microbial culture,of which 7 patients were positive for RPA.The results of RPA amplification from CSF samples of 14 patients were positive.After purification of RPA amplification products from CSF samples of 10 patients,nucleic acid sequences were compared with the target fragment of LepA gene of Streptococcus pneumoniae,and the results were consistent.Conclusion RPA assay can be used for the rapid detection of pneumococcal meningitis.
作者
侯玉婷
俞梦楚
王妍柏
张庆
HOU Yuting;YU Mengchu;WANG Yanbai;ZHANG Qing(Department of Cerebrospinal Fluid Laboratory of Neurology;Institute of Medical Sciences;Department of Neurology,the General Hospital of Ningxia Medical University,Yinchuan,Ningxia 750004,China)
出处
《检验医学与临床》
CAS
2023年第16期2316-2319,共4页
Laboratory Medicine and Clinic
基金
宁夏自然科学基金项目(2020AAC03374)
宁夏医科大学优势学科群建设项目(XY201714)。
关键词
细菌性脑膜炎
肺炎链球菌
重组酶聚合酶扩增技术
脑脊液
bacterial meningitis
Streptococcus pneumoniae
recombinase polymerase amplification assay
cerebrospinal fluid
