Garaizar J, López-Molina N, Laconcha I, Lau Baggesen D, Rementeria A, Vivanco A, Audicana A, Perales I
Department of Immunology, Microbiology and Parasitology, Basque Country University, 01080 Vitoria-Gasteiz, Spain.
Appl Environ Microbiol. 2000 Dec;66(12):5273-81. doi: 10.1128/AEM.66.12.5273-5281.2000.
Strains of Salmonella enterica (n = 212) of different serovars and phage types were used to establish a library typing computerized system for serovar Enteritidis on the basis of PCR fingerprinting, infrequent-restriction-site PCR (IRS-PCR), or pulsed-field gel electrophoresis (PFGE). The rate of PCR fingerprinting interassay and intercenter reproducibility was low and was only increased when DNA samples were extracted at the same time and amplified with the same reaction mixtures. Reproducibility of IRS-PCR technique reached 100%, but discrimination was low (D = 0.52). The PFGE procedure showed an intercenter reproducibility value of 93.3%. The high reproducibility of PFGE combined with the previously determined high discrimination directed its use for library typing. The use of PFGE with enzymes XbaI, BlnI, and SpeI for library typing of serovar Enteritidis was assessed with GelCompar 4.0 software. Three computer libraries of PFGE DNA profiles were constructed, and their ability to recognize new DNA profiles was analyzed. The results obtained pointed out that the combination of PFGE with computerized analysis could be suitable in long-term epidemiological comparison and surveillance of Salmonella serovar Enteritidis, specially if the prevalence of genetic events that could be responsible for changes in PFGE profiles in this serovar was low.
使用不同血清型和噬菌体类型的肠炎沙门氏菌菌株（n = 212），基于聚合酶链反应（PCR）指纹图谱、低频限制性位点PCR（IRS-PCR）或脉冲场凝胶电泳（PFGE），建立了肠炎血清型的文库分型计算机系统。PCR指纹图谱的批间和中心间重复性较低，只有当同时提取DNA样本并用相同反应混合物进行扩增时，重复性才会提高。IRS-PCR技术的重复性达到100%，但鉴别力较低（D = 0.52）。PFGE程序显示中心间重复性值为93.3%。PFGE的高重复性与先前确定的高鉴别力相结合，使其可用于文库分型。使用GelCompar 4.0软件评估了用XbaI、BlnI和SpeI酶进行PFGE对肠炎血清型进行文库分型的情况。构建了三个PFGE DNA图谱的计算机文库，并分析了它们识别新DNA图谱的能力。所得结果指出，PFGE与计算机分析相结合适用于肠炎沙门氏菌血清型的长期流行病学比较和监测，特别是在该血清型中可能导致PFGE图谱变化的遗传事件发生率较低的情况下。
