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通过重复元件PCR对巴尔通体菌种进行基因组指纹分析以区分菌种和菌株。

Genomic fingerprinting of Bartonella species by repetitive element PCR for distinguishing species and isolates.

作者信息

Rodriguez-Barradas M C, Hamill R J, Houston E D, Georghiou P R, Clarridge J E, Regnery R L, Koehler J E

机构信息

Section of Infectious Diseases, Medical Service, Veterans Affairs Medical Center, Houston, TX 77030, USA.

出版信息

J Clin Microbiol. 1995 May;33(5):1089-93. doi: 10.1128/jcm.33.5.1089-1093.1995.

Abstract

Repetitive-element PCR (rep-PCR) with primers based on repetitive extragenic palindromic (REP) and enterobacterial repetitive intergenic consensus (ERIC) repeated DNA sequences was used for genomic finger-printing of Bartonella species. This technique was applied by using either extracted genomic DNA or preparations of whole bacterial cells directly. PCR fingerprints with either the REP-based primers (REP-PCR) or primers based on the ERIC repeat (ERIC-PCR) revealed species-specific band patterns for the various Bartonella isolates. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. ERIC-PCR banding patterns were less complex than those obtained by REP-PCR but allowed better discrimination between strains within species. By combining results of REP-PCR and ERIC-PCR, five different fingerprint profiles were identified among 17 isolates of Bartonella henselae, but only one profile was identified among the five isolates of Bartonella quintana. Other Bartonella species yielded distinct rep-PCR fingerprints. rep-PCR is a useful technique for identification of Bartonella organisms to the species level and offers the advantage of ease of performance, with only small quantities of cells needed for the whole-cell procedure. This technique also appears to be useful for subtyping B. henselae isolates.

摘要

使用基于重复基因外回文序列(REP)和肠杆菌重复基因间共有序列(ERIC)重复DNA序列的引物进行重复元件PCR(rep-PCR),用于巴尔通体菌种的基因组指纹分析。该技术可通过使用提取的基因组DNA或直接使用全细菌细胞制剂来应用。使用基于REP的引物(REP-PCR)或基于ERIC重复序列的引物(ERIC-PCR)进行PCR指纹分析,可揭示各种巴尔通体分离株的种特异性条带模式。从提取的基因组DNA的rep-PCR或全细胞制剂获得的DNA指纹图谱产生了可比的模式。ERIC-PCR条带模式比REP-PCR获得的条带模式更简单,但能更好地区分种内菌株。通过结合REP-PCR和ERIC-PCR的结果,在17株亨氏巴尔通体分离株中鉴定出五种不同的指纹图谱,但在五日热巴尔通体的五株分离株中仅鉴定出一种图谱。其他巴尔通体菌种产生了独特的rep-PCR指纹图谱。rep-PCR是一种用于将巴尔通体菌鉴定到种水平的有用技术,具有操作简便的优点,全细胞程序仅需少量细胞。该技术似乎也可用于亨氏巴尔通体分离株的亚型分析。

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