Unal S, Hoskins J, Flokowitsch J E, Wu C Y, Preston D A, Skatrud P L
Infectious Disease Research, Eli Lilly and Co., Indianapolis, Indiana 46285.
J Clin Microbiol. 1992 Jul;30(7):1685-91. doi: 10.1128/jcm.30.7.1685-1691.1992.
A polymerase chain reaction (PCR)-based test was developed for the detection of mecA in staphylococci. To facilitate this process, a rapid cell lysis procedure was established for the release of DNA from staphylococcal strains. Primers based on the DNA sequence of the mecA gene from Staphylococcus aureus were used in PCRs to screen for the presence of this gene in a total of 98 staphylococcal isolates. Fifty-one isolates were mecA positive (17 S. aureus strains and 34 coagulase-negative staphylococci including S. epidermidis, S. haemolyticus, and S. simulans). Results obtained with PCRs were generally consistent with those of standard microbiological assays. PCRs designed to detect the femA gene (factor essential for methicillin resistance) revealed the presence of the gene in all S. aureus strains examined regardless of the susceptibility profiles of the strains to methicillin. In contrast, femA could not be detected in coagulase-negative staphylococci by PCR with the same primers. Low-stringency hybridization suggested the presence of a gene structurally related to femA in S. epidermidis and other coagulase-negative staphylococci examined.
开发了一种基于聚合酶链反应(PCR)的检测葡萄球菌中mecA基因的试验。为便于此过程,建立了一种快速细胞裂解程序以从葡萄球菌菌株中释放DNA。在PCR中使用基于金黄色葡萄球菌mecA基因DNA序列的引物,以筛选98株葡萄球菌分离株中该基因的存在情况。51株分离株mecA呈阳性(17株金黄色葡萄球菌菌株和34株凝固酶阴性葡萄球菌,包括表皮葡萄球菌、溶血葡萄球菌和模仿葡萄球菌)。PCR获得的结果通常与标准微生物学检测结果一致。设计用于检测femA基因(耐甲氧西林必需因子)的PCR显示,在所检测的所有金黄色葡萄球菌菌株中均存在该基因,无论这些菌株对甲氧西林的敏感性如何。相比之下,使用相同引物通过PCR在凝固酶阴性葡萄球菌中未检测到femA。低严谨度杂交表明,在所检测的表皮葡萄球菌和其他凝固酶阴性葡萄球菌中存在与femA结构相关的基因。