Profiling the Age-related Microbiome via Detection of Antibodies to Gut Bacteria

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Asy Syifa Labibah(Asy Syifa Labibah) 查詢紙本館藏

畢業系所

生醫科學與工程學系

論文名稱

Profiling the Age-related Microbiome via Detection of Antibodies to Gut Bacteria
(Profiling the Age-related Microbiome via Detection of Antibodies to Gut Bacteria)

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摘要(中)

德氏乳桿菌（L. delbreuckii）是一種益生菌，可與嗜熱鏈球菌一起存在於優酪乳中。之前的研究發現，攝入用 (L. delbreuckii) 發酵的優酪乳可以提高有助於老年人生物防禦的免疫系統參數值。大腸桿菌（E.coli）是人類腸道微生物組中的一種機會性細菌。這種細菌會引起腸胃炎和腸道感染。與年輕人相比，在老年人中發現了不同的腸道微生物群組成，其中某些病原體增加。這可能與老年人免疫能力下降有關。在這項研究中，我們通過檢測腸道細菌的抗體來分析不同年齡段的差異腸道微生物組。側向流動免疫分析 (LFIA) 是一種快速且低成本的方法，用於檢測人血清和唾液中的腸道細菌抗體。 L. delbreuckii 或 E.coli 的抗體可以通過 LFIA 在人血清和唾液中檢測到，該 LFIA 用細菌裂解物點在硝酸纖維素膜上。此外，老年人的大腸桿菌或德氏乳桿菌的抗體高於年輕人。在老年人中，大腸桿菌的抗體比德氏乳桿菌的抗體高得多。另一方面，德氏乳桿菌產生的電力明顯高於大腸桿菌。分析不同年齡段腸道中生電細菌的抗體可能會開啟一項關於衰老的新研究。

摘要(英)

Abstract

Lactobacillus delbrueckii (L. delbreuckii) is one of probiotic bacteria that can be in the yogurt with Streptococcus thermophilus bacteria. The previous study has found that the intake of yogurt fermented with (L. delbreuckii) can improve the values of immune system parameters that contributed to biological defense in the elderly. Eschericia coli (E. coli) is an opportunistic bacterium in the human gut microbiome. This bacterium can cause gastroenteritis and intestinal infection. In older people, a different gut microbiota composition with the increased certain pathogens was found, as compared to younger people. This may be related to a decrease in immunocompetence in elder people. In this study, we profile the differential gut microbiome in various ages via detection of antibodies to gut bacteria. The lateral flow immunoassay (LFIA), a rapid and low cost method, was constructed to detect the antibodies to gut bacteria in the human sera and saliva. The antibodies to L. delbreuckii or E. coli can be detected in the human sera and saliva by LFIA spotted with bacterial lysates on nitrocellulose membrane. Furthermore, the antibodies to E. coli or L. delbreuckii were higher on older people than young people. In older people, the antibodies to E. coli were much higher than antibodies to L.delbrueckii. On the other hand, electricity produced by L. delbrueckii was significantly higher than that by E. coli. Profiling the antibodies to electrogenic bacteria in the gut of various ages may open a new study of aging.

Keyword : L. delbrueckii,E. coli, LFIA, Aging factor

關鍵字(中)

★ 大腸桿菌
★ 老化因數

關鍵字(英)

★ L. delbrueckii
★ E. coli
★ LFIA
★ Aging factor

論文目次

Table of Contents

Abstract …………………………………………………………………………….. vii
Acknowledgments ………………………………………………………………….. viii
Table of Contents …………………………………………………………………… ix
List of Figures ………………………………………………………………………. x
List of Table ………………………………………………………………………… xi
Abbreviation List ……………………………………………………………………. xii
Chapter 1 Introduction ……………………………………………………………… 1
Chapter 2 Literature review …………………………………………………………. 3
2.1 Gut Bacteria …………………………………………………………... 3
2.1.1 Lactobacillus delbrueckii ………………………………………... 4
2.1.2 Escherichia coli ………………………………………………… 5
2.2 Elderly Gut ……………………………………………………………. 6
2.3 LFIA ………………………………………………………………….. 8
2.4 Electrogenic Bacteria …………………………………………………. 12
2.5 MFC …………………………………………………………………… 14
Chapter 3 Materials and Methods …………………………………………………… 16
3.1 Ethics Statement ……………………………………………………….. 16
3.2 Identification bacteria ………………………………………………… 16
3.3 Bacterial Culture and Bacterial Lysate ……………………………….. 16
3.4 LFIA Fabrication ……………………………………………………… 17
3.5 Extraction DNA fecal and RT-qPCR …………………………………. 17
3.6 Immunization and Titer Antibody …………………………………….. 18
3.7 Electricity detection in vitro …………………………………………… 19
3.8 Statistics analysis …………………………………………………… ... 19
Chapter 4 Result …………………………………………………………………… . 20
4.1 Screening L. delbrueckii ssp. Bulgaricus bacteria from human fecal sample ………………………………………………………………… 20
4.2 Determine antibodies against L.delbrueckii and E.coli in humans
and mice ……………………………………………………………… 24
4.3 Determine antibodies against L. delbrueckii and E. coli in different age human and mice …………………………………………………….. . 25
4.4. Immunization Result and Antibody Titer ………………………….. . 27
4.5. Electricity production of gut bacteria ……………………………….. 29
Chapter 5 Discussion and Conclusion ………………………………………............ 31
Reference ……………………………………………………………………………. 35

參考文獻

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指導教授

黃俊銘(Eric)

審核日期

2021-11-1

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