Sekiguchi K, Saito M, Yajima R
Dainabot Co., Ltd., Research Center, Chiba, Japan.
Microbiol Immunol. 1995;39(8):545-50. doi: 10.1111/j.1348-0421.1995.tb02240.x.
Ten kinds of peptides (21 to 32 amino acids in length) were synthesized based on the reported amino acid sequences of the penicillin-binding protein 2' (PBP2') of methicillin-resistant Staphylococcus aureus (MRSA). Antibodies against these synthetic peptides (SPs) were generated by immunizing rabbits. The antibodies raised against all the peptides except for one reacted to PBP2' of MRSA and to SPs used for immunization but not to any other protein of MRSA or methicillin-susceptible S. aureus (MSSA) tested by ELISA and Western blotting. A sandwich immunoradiometric assay (IRMA) for the detection of PBP2' was developed using these antibodies. The method could detect PBP2' extracted from as few as 3 x 10(4) cells of a clinical MRSA isolate, and a good correlation between cell number and signal radio-count was observed. IRMA was positive for all 51 methicillin-resistant staphylococci isolated from patients, and was negative for all the 28 methicillin-susceptible ones and 19 strains of other bacterial species. IRMA could be a simple and reliable method for MRSA detection in the clinical bacterial laboratory.
基于耐甲氧西林金黄色葡萄球菌（MRSA）青霉素结合蛋白2'（PBP2'）已报道的氨基酸序列，合成了10种肽（长度为21至32个氨基酸）。通过免疫兔子产生针对这些合成肽（SPs）的抗体。除一种肽外，针对所有肽产生的抗体通过ELISA和蛋白质印迹法检测，与MRSA 的PBP2'以及用于免疫的SPs发生反应，但不与MRSA或甲氧西林敏感金黄色葡萄球菌（MSSA）的任何其他蛋白质发生反应。利用这些抗体开发了一种用于检测PBP2'的夹心免疫放射分析（IRMA）方法。该方法能够检测从临床MRSA分离株中少至3×10⁴个细胞中提取的PBP2'，并且观察到细胞数量与信号放射计数之间具有良好的相关性。IRMA对从患者中分离出的所有51株耐甲氧西林葡萄球菌呈阳性，对所有28株甲氧西林敏感葡萄球菌和19株其他细菌菌株呈阴性。IRMA可能是临床细菌实验室中检测MRSA的一种简单可靠的方法。
