为建立一种有效、快速、低成本区分长双歧杆菌婴儿亚种的方法,对13株基因组测定的长双歧杆菌构建了基于同源基因的系统进化树,确认其中7株为长亚种,5株为婴儿亚种,1株为猪亚种。通过碳水化合物利用谱比较和特异性基因扩增的手段确认可有效用于快速区分长双歧杆菌婴儿亚种的方法。结果显示,同一亚种的菌株利用同一种碳水化合物的能力存在差异,表明无法根据碳水化合物的利用来区分长双歧杆菌的不同亚种;长双歧杆菌长亚种糖激酶基因和长双歧杆菌婴儿亚种唾液酸酶基因可有效用于长双歧杆菌婴儿亚种的快速区分,并用该方法对40株长双歧杆菌进行了亚种确认,其中5株为长双歧杆菌婴儿亚种。后随机选择了其中的4株菌进行基因组草图测序,经系统进化树构建,确认为长双歧杆菌婴儿亚种,并进一步证实特异性基因扩增可实现长双歧杆菌婴儿亚种的快速区分。
The aim of the current research was to establish an effective, rapid and low-cost method to identify Bifidobacterium longum subsp. infantis. The subspecies of 13 genomes sequenced B. longum strains were confirmed according to the phylogenetic tree based on orthologous genes in 13 subspecies, seven strains were B. longum subsp. longum, five strains were B. longum subsp. infantis and one was B. longum subsp. suis. A method for rapidly distinguishing B. longum subsp. infantis was confirmed by analyzing their carbohydrate utilization capacity and amplification of specific genes. The results indicated that it was impossible to distinguish subspecies based on carbohydrate utilization capacity due to strains from each subspecies were different. As comparison, the sugar kinase genes in B. longum subsp. longum and sialidase genes in B. longum subsp. infantis could be efficiently used for distinguishing B. longum subsp. infantis. With this method, five strains of B. longum subsp. infantis were identified from 40 strains of B. longum. Four of the five strains of B. longum subsp. infantis were randomly selected and further confirmed as B. longum subsp. infantis. Therefore, this study verifies that B. longum subsp. infantis can be rapidly distinguished by amplifying specific genes.
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