应用基于 PCR 的技术鉴定越南腹泻儿童粪便样本中优势细菌的遗传指纹多样性。

IF 3.4 Q2 INFECTIOUS DISEASES
Thi Quy Nguyen, Trong Khoa Dao, Hong Duong Nguyen, Thi Bich Thuy Phung, Thi Thanh Nga Pham, Thi Viet Ha Nguyen, Thi Huong Trinh, Huu Cuong Le, Thi Thu Hong Le, Thi Huyen Do
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引用次数: 0

摘要

在越南,腹泻(尤其是顽固性腹泻)是儿童最常见的疾病之一,而相当一部分病例的病原体呈阴性;因此,迫切需要了解肠道细菌失调的情况。本研究将 5 名健康儿童和 10 名腹泻儿童粪便样本中的细菌与其他残留物分离,然后采用 PCR-RFLP 和 PCR-DGGE 方法提取其元基因组 DNA,根据 V3、V6-V8 区域和 16S rRNA 基因评估其多样性。结果,使用 GeneJET 试剂盒和化学方法成功提取了高质量、高数量和高多样性的细菌元基因组 DNA。对凝胶中具有代表性的 73 个 DNA 片段进行的序列分析表明,所有腹泻组均存在明显的细菌菌群失调。病毒性腹泻的特点是细菌多样性极度减少,双歧杆菌和链球菌遍地开花。在持续性腹泻中,链球菌的数量也最多。双歧杆菌、乳酸杆菌和肠球菌等可能在肠道细菌群落的自我平衡中发挥作用的有益细菌在所有腹泻组中都能看到,而乳酸杆菌和 Akkermansia muciniphila 在健康组中能看到,但在腹泻组中却看不到。这项研究为肠道细菌失调与儿童腹泻之间的关系提供了更多证据,强调了链球菌的增加。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Application of PCR-Based Techniques for the Identification of Genetic Fingerprint Diversity of Dominant Bacteria in Fecal Samples of Children with Diarrhea in Vietnam.

In Vietnam, diarrhea, especially persistent diarrhea, is one of the most common diseases in children, while a significant proportion of cases are negative with pathogens; thus, there is an urgent need to understand gut bacterial dysbiosis. In this study, bacteria in the fecal samples of five healthy and ten diarrheal children were separated from other residues, then adopted to extract their metagenomic DNA for evaluating their diversity based on V3 and V6-V8 regions and the 16S rRNA gene by PCR-RFLP and PCR-DGGE. As a result, bacterial metagenomic DNAs with high quality, quantity and diversity were successfully extracted using a GeneJET kit and a chemical protocol. A sequence analysis of 73 representative DNA fragments from gels indicated a remarkable bacterial dysbiosis in all groups of diarrhea. Viral diarrhea was characterized by extremely reduced bacterial diversity with the blossom of Bifidobacterium and Streptococcus. Streptococcus was also the most abundant in persistent diarrhea. Beneficial bacteria that may play a role in the self- rebalance in intestinal bacterial communities, such as Bifidobacterium, Lactobacillus, and Enterococcus, were seen in all diarrheal groups, while Bacteroides and Akkermansia muciniphila were seen in the healthy group but absent in the diarrheal groups. This study provides additional evidence for a relationship between intestinal bacterial dysbiosis and diarrhea in children, emphasizing an increase in Streptococcus.

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来源期刊
Infectious Disease Reports
Infectious Disease Reports INFECTIOUS DISEASES-
CiteScore
5.10
自引率
0.00%
发文量
82
审稿时长
11 weeks
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