https://scholars.lib.ntu.edu.tw/handle/123456789/146079
標題: | Determination of Enterococcus Faecalis Groesl Full-Length Sequence and Application for Species Identification | 作者: | TENG, LEE-JENE HSUEH, PO-REN WANG, YI- HUI LIN, HSIAO-MANN LUH, KWEN-TAY HO, SHEN-WU |
公開日期: | 2001 | 卷: | v.39 | 期: | n.9 | 起(迄)頁: | 3326-3331 | 來源出版物: | JOURNAL OF CLINICAL MICROBIOLOGY | 摘要: | Amplification of the partial Cpn60 (or GroEL) gene segment has been used for identification of many bacteria including Enterococcus species. To obtain more sequence data from groESL genes of Enterococcus faecalis, the full-length sequence of the E. faecalis groESL genes containing groES ( 285 bp), spacer (57 bp), and groEL (1626 bp) was determined. A database search from GenBank revealed that the deduced E. faecalis GroES and GroEL proteins show significant homology to the GroES and GroEL proteins of other bacteria. The GroEL (groEL) of E. faecalis had the highest identity with Streptococcus pneumoniae (81.8% amino acid sequence identity and 73.0% nucleotide sequence identity), followed by Lactococcus zeae, while GroES (groES) had 60.2% (64.6%) identity with Lactobacillus zeae, and 58.5% (66.2%) with Lactococcus lactis, followed by 57.0% (65.5%) with Bacillus subtilis. Based on the groES sequence, an E. faecalis- specific PCR assay was developed and this PCR assay was positive for all the E. faecalis strains tested. Dot blot hybridization using either groES or groEL as probe distinguished E. faecalis clearly from other species, indicating that both genes can be used as suitable targets for E. faecalis identification. Moreover, broad-range PCR- RFLP of groESL was designed to differentiate eight commonly encountered Enterococcus species. The Enterococcus species of reference strains could be easily differentiated on the basis of restriction patterns produced by HaeIII and RsaI. The DNA- based assays developed in this study provide an alternative to currently used methods of identification for clinically important enterococcal species. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/89825 |
顯示於: | 醫學檢驗暨生物技術學系 |
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