Parra Eliana, Castañeda Elizabeth, Moreno Jaime
Grupo de Microbiología, Instituto Nacional de Salud, Bogotá, DC, Colombia.
Biomedica. 2007 Sep;27(3):454-60.
Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis are the main human pathogens that cause meningitis.
Primers omp2, lytA and crgA were evaluated with H. influenzae, S. pnumoniae and N. meningitidis DNA in a multiplex PCR, determining the sensitivity and the specificity of the technique.
Primers for H. influenzae outer membrane protein (omp2, 1000 pb), S. pneumoniae autolysin (lytA, 395 pb) and N. meningitidis, contact regulated gene (crgA, 230 pb) were evaluated in a multiplex PCR, determining the sensitivity and the specificity of the technique.
Reproducible results were obtained with 50 nM of each of the three primers and annealing temperature of 57 degrees C in the multiplex PCR, obtaining a sensitivity of 12.5 fg for H. influenzae and S. pneumoniae and 3.12 fg for N. meningitidis. No cross reactions with other microorganisms agents of meningitis or related with the genera, appeared.
The results for sensitivity and specificity suggest that the evaluated primers can be used for the development of a PCR in a multiplex format to the identification of the three main pathogens that cause meningitis.
流感嗜血杆菌、肺炎链球菌和脑膜炎奈瑟菌是引起脑膜炎的主要人类病原体。
在多重聚合酶链反应（PCR）中，用流感嗜血杆菌、肺炎链球菌和脑膜炎奈瑟菌的DNA评估引物omp2、lytA和crgA，确定该技术的敏感性和特异性。
在多重PCR中评估流感嗜血杆菌外膜蛋白（omp2，1000 pb）、肺炎链球菌自溶素（lytA，395 pb）和脑膜炎奈瑟菌接触调节基因（crgA，230 pb）的引物，确定该技术的敏感性和特异性。
在多重PCR中，三种引物各50 nM且退火温度为57℃时获得了可重复的结果，流感嗜血杆菌和肺炎链球菌的敏感性为12.5 fg，脑膜炎奈瑟菌的敏感性为3.12 fg。未出现与其他脑膜炎微生物病原体或相关属的交叉反应。
敏感性和特异性结果表明，所评估的引物可用于开发多重PCR，以鉴定引起脑膜炎的三种主要病原体。
