N­-糖苷酶(PNGase)是真核生物中的常见酶，但在原核生物中极为罕见，目前仅报道在脑膜炎败血伊丽莎白金菌(EM)中存在。本文旨在检测PNGase基因在EM临床分离株中的分布，为该菌的亚型分类及快速核酸分子检测提供依据，为研究PNGase在原核生物中的生物学功能奠定基础。于2010年7月～2012年12月收集浙江省3家三级甲等医院的65株EM临床分离株，提取细菌基因组DNA，利用16S rRNA细菌通用引物、PNGase F 和PNGase F­Ⅱ 特异性引物，进行组合聚合酶链反应(PCR)。阳性扩增产物测序后与美国国立生物技术信息中心(NCBI)数据库中已知EM序列比对确认。结果显示，PNGase阳性菌64株，总阳性率为98.5%；PNGase F和PNGase F­Ⅱ共阳性菌39株，共阳性率为60.0%。PNGase F阳性菌41株，阳性率为63.1%；PNGase F­Ⅱ阳性菌62株，阳性率为95.4%，PNGase F­Ⅱ阳性率高于PNGase F (配对χ²检验，P<0.01)，提示PNGase可能对EM具有重要的生物学意义。结果还表明，本研究建立的组合PCR可应用于EM菌株中PNGase基因分型。

The present paper aims to investigate the distribution of two peptide: N­-glycanase (PNGase) genes in Elizabethkingia meningosepticum (EM). The genomic DNAs of 65 clinical strains isolated from 3 hospitals in Zhejiang Province were extracted and subjected to polymerase chain reaction (PCR) for 16S rRNA, PNGase F and PNGase F­-Ⅱ genes respectively. The PCR products were sequenced and compared to the known sequences in National Center of Biotechnology Information (NCBI). The results showed that the positive rate of PNGase gene was 98.5% and the PNGase F/PNGase F­-Ⅱ gene co­existence rate was 60.0%. The positive rates of PNGase F gene and PNGase F­-Ⅱ gene were 63.1% and 95.4%, respectively (matching χ² test, P<0.01). The results indicate that the developed PCR can be used to detect PNGase gene in EM strains and PNGase activity may be crucial for the survival.