摘要
目的建立快速、高通量的多重PCR/反向线点杂交(RLB)的方法检测3种细菌性脑膜炎病原体--肺炎链球菌、流感嗜血杆菌和脑膜炎奈瑟菌。方法针对肺炎链球菌、流感嗜血杆菌和脑膜炎奈瑟菌的特异性基因序列设计引物和相应的特异性探针,优化多重PCR/RLB检测中的引物浓度、dNTP浓度、探针的浓度等的条件。结果多重PCR/RLB检测对肺炎链球菌DNA最低检测极限可达6.2×10-1ng/μl,对流感嗜血杆菌DNA最低检测极限可达8.5×10-8ng/μl,对脑膜炎奈瑟菌DNA最低检测极限可达8.3×10-8ng/μl。结论多重PCR/RLB检测体系能够正确的鉴定3种细菌性脑膜炎病原体,用于临床样本的高通量筛查细菌性脑膜炎病原体,特异性好,灵敏度高。
Objective To develop a rapid,high-throughput screening method of a multiplex PCR-based reverse line blot（mPCR/RLB） array to detect S.pneumoniae,H.influenzae and N.meningitides.Methods Highly validated specific primers and probes were used to diagnostic regions unique to each pathogen.Primers,dNTP,and probes in various concentrations were detected to determine the optimal conditions.Results The limit of detection of mPCR/RLB array was 6.2×10-1ng/μl for S.pneumoniae,with 8.5×10-8ng/μl genome DNA for H.influenzae,and 8.3×10-8ng/μl for N.meningitides.Conclutions mPCR/RLB array can detect three pathogen correctly,and it is valuable for high-throughput screening pathogens of clinical samples of bacterial meningitis.
出处
《实验与检验医学》
CAS
2011年第4期347-349,共3页
Experimental and Laboratory Medicine
基金
深圳市科技计划资助项目(200902040)
