van Belkum A, Bax R, Peerbooms P, Goessens W H, van Leeuwen N, Quint W G
Diagnostic Centre SSDZ, Delft, The Netherlands.
J Clin Microbiol. 1993 Apr;31(4):798-803. doi: 10.1128/jcm.31.4.798-803.1993.
A typing procedure for methicillin-resistant Staphylococcus aureus (MRSA) based on the polymerase chain reaction (PCR) amplification of both mecA sequences and variable DNA sequences as present in the prokaryotic genome has been developed. Two primers based on the sequences of DNA repeats as discovered in gram-negative members of the family Enterobacteriaceae allow detection of variable regions in the genome of a gram-positive bacterium such as S. aureus, as does a newly described arbitrary primer. This procedure, enabling the detection of 23 different genotypes in a collection of 48 MRSA isolates, was validated by comparisons with phage typing studies. It appeared that within the same group of isolates only 13 different phagovars could be identified. Combination of the results from both phage typing and genotyping allowed the discrimination of 34 of 48 isolates. However, depending on the primer-variable complexity of the PCR fingerprints, which could also be modulated by combination of PCR primers, clear homologies between the groups defined by either phage typing or fingerprinting were observed. An analysis of an MRSA outbreak in a geriatric institution showed a collection of genetically homogeneous isolates. In agreement with phage typing, PCR fingerprinting revealed the identical natures of the MRSA strains isolated from all patients.
已开发出一种基于聚合酶链反应（PCR）扩增mecA序列和原核基因组中存在的可变DNA序列的耐甲氧西林金黄色葡萄球菌（MRSA）分型方法。基于在肠杆菌科革兰氏阴性菌成员中发现的DNA重复序列的两个引物，能够检测革兰氏阳性菌如金黄色葡萄球菌基因组中的可变区域，一种新描述的任意引物也有此功能。通过与噬菌体分型研究进行比较，验证了该方法能够检测48株MRSA分离株中的23种不同基因型。结果表明，在同一组分离株中只能鉴定出13种不同的噬菌体型。噬菌体分型和基因分型的结果相结合，可区分48株分离株中的34株。然而，根据PCR指纹图谱的引物可变复杂性（也可通过PCR引物组合进行调节），观察到噬菌体分型或指纹图谱定义的组之间存在明显的同源性。对一家老年机构中MRSA暴发的分析显示，分离株在基因上具有同质性。与噬菌体分型结果一致，PCR指纹图谱显示从所有患者分离出的MRSA菌株具有相同的性质。
