Benito M J, Rodríguez M M, Córdoba M G, Aranda E, Córdoba J J
Higiene de los Alimentos, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
Lett Appl Microbiol. 2000 Nov;31(5):368-73. doi: 10.1046/j.1472-765x.2000.00833.x.
An arbitrarily primed-polymerase chain reaction (AP-PCR) method was optimized to differentiate Staphylococcus aureus from other staphylococcal species, using DNA from crude cell extract. From the different assays carried out, the best resolution of the band patterns was obtained when the reaction mixture contained 200 micromol l(-1) dNTPs, 200 ng primer, 1 U Taq DNA polymerase and 3 mmol l(-1) MgCl2 and the amplification conditions were: initial denaturation of 94 degrees C for 1 min, primer annealing of 30 degrees C for 1.5 min, DNA extension at 55 degrees C for 5 min and final extension at 55 degrees C for 5 min. The results of the characterization of the staphylococcal isolates by AP-PCR are in accordance with those of the biochemical identification by the API Staph System, time of analysis of the AP-PCR being only 6-7 h. Thus, this technique could be a useful method for microbial quality assurance.
优化了一种任意引物聚合酶链反应(AP-PCR)方法,以利用粗细胞提取物中的DNA区分金黄色葡萄球菌与其他葡萄球菌属菌种。在进行的不同试验中,当反应混合物含有200微摩尔/升dNTP、200纳克引物、1单位Taq DNA聚合酶和3毫摩尔/升氯化镁,且扩增条件为:94℃初始变性1分钟、30℃引物退火1.5分钟、55℃DNA延伸5分钟以及55℃最终延伸5分钟时,可获得最佳的条带模式分辨率。通过AP-PCR对葡萄球菌分离株进行鉴定的结果与使用API葡萄球菌系统进行生化鉴定的结果一致,AP-PCR的分析时间仅为6 - 7小时。因此,该技术可能是一种用于微生物质量保证的有用方法。