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翟刚.16S-rRNA测序技术分析早产儿肠道细菌基因组指导新生儿坏死性小肠结肠炎手术时机选择的研究[J].中国医药科学,2024,14(11):130-133        基金项目:广西壮族自治区卫生健康委员会自筹经费科研课题(Z20200951)
16S-rRNA测序技术分析早产儿肠道细菌基因组指导新生儿坏死性小肠结肠炎手术时机选择的研究
16S-rRNA sequencing technology to analyze the intestinal bacterial genome of premature infants to guide the timing of NEC surgery
  
DOI:
中文关键词:  新生儿坏死性小肠结肠炎;16S rRNA测序技术;肠道细菌基因;手术时机
英文关键词:Necrotizing enterocolitis of newborn; 16S-rRNA sequencing technology; Intestinal bacterial gene; Timing of operation
作者单位
翟刚 广西壮族自治区百色市人民医院胃肠外科,广西百色 533000 
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中文摘要:
      [摘要] 目的 探讨16S-rRNA测序技术分析早产儿肠道细菌基因组指导新生儿坏死性小肠结肠炎(NEC)手术时机选择。 方法 前瞻性选择2021年1月至2022年6月百色市人民医院需要手术治疗的NEC患儿30例为观察组,选择同期内科保守治疗的Ⅰ期15例和Ⅱa期15例患者为对照组。采用HiSeq测序平台,借助双端测序模式进行高通量二代测序,比较两组多样性指数、优势均属丰度及不同优势菌比值等;绘制受试者操作特征(ROC)曲线,分析16S-rRNA测序技术的指导价值。 结果 60例患者60份样本中共获得细菌84个,且两组样品均为副杆状菌属最高,其次为Ruminococcus、Blautia、Aeromonas和Fusobacterium;两组肠道菌群上述菌门丰度存在差异(P < 0.05);从粪便标本中共获得有效序列7347 481条,人均130 857条,测序平均覆盖度为(92.15±5.61)%;观察组手术治疗的NEC患儿中香农-维纳(Shannon)及辛普森多样性(Simpson)指数低于对照组内科保守治疗患儿,差异有统计学意义(P < 0.05);ROC曲线结果表明,16S-rRNA测序技术在NEC患儿手术时机选择中的指导AUC为0.846,指导灵敏度为87.51%,特异度为83.16%。 结论 NEC患儿常伴有肠道细菌基因组改变,且菌群结构的变化与患儿病情严重程度有关,通过16S-rRNA测序技术能指导NEC患儿手术治疗时机。
英文摘要:
      [Abstract] Objective To investigate the 16S-rRNA sequencing technology to analyze the intestinal bacterial genome of premature infants to guide the timing of necrotizing enterocolitis of newborn (NEC) surgery. Methods Cases of pediatric patients with NEC admitted to Baise People’s Hospital from January 2021 to June 2022 were prospectively selected and divided into the observation group (n=30) and the control group (n=30). The observation group was required to be treated with surgery, while the control group, including 15 cases in phase Ⅰ and 15 case in phase Ⅱa, was conservatively treated in the Department of Internal Medicine during the same period. HiSeq sequencing platform was used, high-throughput second-generation sequencing was carried out with the help of double-end sequencing mode. The diversity index, abundance of dominant species and ratio of different dominant bacteria were compared between the two groups. The receiver operating characteristic (ROC) curves were drawn and the guiding value of 16S-rRNA sequencing technology was analyzed. Results A total of 84 bacteria were obtained from 60 samples of 60 patients, and Parabacteroides was the highest in both groups of species, followed by Ruminococcus, Blautia, Aeromonas and Fusobacterium. There was statistically significant difference in the abundance of the above phylum between the two groups of gut microbiota (P < 0.05). A total of 7347 481 valid sequences were obtained from fecal specimens, with an average of 130 857 sequences per capita, and the average sequencing coverage was (92.15±5.61)%. The Shannon-wiener and Simpson diversity index in pediatric patients with NEC treated with surgery in the observation group was lower than that in the control group treated with conservative internal medicine, with statistically significant difference (P < 0.05). The ROC curve results showed that the guidance AUC of 16S-rRNA sequencing technology in the selection of surgical timing for pediatric patients with NEC was 0.846, the guidance sensitivity was 87.51%, and the specificity was 83.16%. Conclusion Pediatric patients with NEC are often accompanied by changes in the genome of intestinal bacteria, and the changes in the flora structure are related to the severity of their illness. 16S-rRNA sequencing technology can guide the timing of surgical treatment for pediatric patients with NEC.
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