Kesanopoulos Konstantinos, Tzanakaki Georgina, Levidiotou Stamatina, Blackwell Caroline, Kremastinou Jenny
National Meningococcal Reference Laboratory, National School of Public Health, 196 Alexandras Avenue, Athens, Greece.
FEMS Immunol Med Microbiol. 2005 Mar 1;43(3):419-24. doi: 10.1016/j.femsim.2004.10.011.
Antibiotic treatment prior to transport or admission of patients to hospital has reduced the proportion of patients with invasive meningococcal disease (IMD) from whom Neisseria meningitidis can be isolated by standard microbiological techniques. Assays to detect the crgA gene were used to detect meningococcal DNA by both conventional polymerase chain reaction (PCR) and real-time PCR (RTPCR) in relation to microbiological diagnosis of cases over two years between 2002 and 2003. The sensitivity of both PCR assays for culture-confirmed cases was 93% and the specificity was 98.6%. Agreement between the two PCR assays was 96.2%. The inter- and intra-assay variations and effects of different amounts of DNA on the melting temperatures were examined. The touch-down RTPCR based on SYBR Green I fluorescent dye detected and characterized N. meningitidis in clinical samples within one hour.
在将患者转运或收治入院之前进行抗生素治疗，已降低了通过标准微生物技术可分离出脑膜炎奈瑟菌的侵袭性脑膜炎球菌病（IMD）患者的比例。在2002年至2003年的两年间，针对病例的微生物学诊断，采用检测crgA基因的检测方法，通过常规聚合酶链反应（PCR）和实时PCR（RTPCR）来检测脑膜炎球菌DNA。两种PCR检测方法对培养确诊病例的灵敏度均为93%，特异性为98.6%。两种PCR检测方法之间的一致性为96.2%。研究了不同检测方法之间及同一检测方法内部的变异情况，以及不同量的DNA对解链温度的影响。基于SYBR Green I荧光染料的降落式RTPCR在一小时内即可检测并鉴定临床样本中的脑膜炎奈瑟菌。
