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发明公开
CN107058564A 儿童自闭症肠道菌群多样性的分析方法及特异性引物对 无效 - 驳回

儿童自闭症肠道菌群多样性的分析方法及特异性引物对
摘要：
本发明提供了一种基于二代高通量测序技术的肠道菌群多样性的分析方法及特异性引物对。其中，该方法包括如下步骤：1)提取待测粪便样本的DNA；2)配置PCR反应体系，进行细菌16S rDNA V1‑V2区的PCR扩增反应；上述PCR反应体系中包含PCR扩增模板、通用引物对等；3)将PCR产物进行目标片段的检测以及PCR产物的纯化；4)将纯化后的PCR产物进行二代高通量测序，检测粪便样本中的微生物的种类；5)利用Illumina MiSeq测序系统进行测序；6)利用FLASH，QIIME，usearch61进行生物信息学分析。本方法通过二代高通量测序技术，对疾病患者肠道菌群多样性及差异性进行检测，与传统菌群检测方法相比，应用新一代高通量技术能够更详细、更深层地解析菌群的组成与功能。
摘要（英）：

The invention provides a method for analyzing children autism intestinal flora diversity and a specific primer pair. The method comprises the following steps: 1) extracting DNA of a to-be-detected stool sample; 2) configuring a PCR reaction system and performing PCR amplified reaction in bacteria 16S rDNA V1-V2 areas, wherein the PCR reaction system comprises a PCR amplification template, a universal primer pair and the like; 3) detecting a target fragment of a PCR product and purifying the PCR product; 4) performing second-generation high-throughput sequencing on the purified PCR product and detecting the types of microorganisms in the stool sample; 5) sequencing by utilizing an I11umina MiSeq sequencing system; 6) performing bio-information analysis by utilizing FLASH, QIIME and usearch61. The method provided by the invention has the benefits that through a second-generation high-throughput sequencing technology, the intestinal flora diversity of a diseased patient and the difference are detected; compared with a traditional flora detection method, the compositions and the functions of floras can be more detailedly and deeply analyzed by applying the new-generation high-throughput technology.

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