Fingerprint analysis of gut microbiota by denaturing gradient gel electrophoresis in newly diagnosed tuberculosis patients

doi:10.3969/j.issn.1000-6621.2019.09.013

Abstract

Abstract:

Objective To analyze the difference in gut microbiota between normal and newly diagnosed tuberculosis patients.Methods From August 2017 to July 2018, 2 to 3 copies of the fecal specimens of healthy medical examinations that met the inclusion criteria were collected at the fixed time every month in the Xi’dian Group Hospital. Specimens collection was stopped after collecting 30 specimens which served as the control group. The fecal specimens of patients with newly diagnosed tuberculosis were collected from August 2017 to July 2018 in the Xi’an Chest Hospital. Similarly, inclusion was stopped once 30 specimens were collected as the tuberculosis group. The diversity and similarity of gut microbiota in the control group and the tuberculosis group were analyzed by denaturing gradient gel electrophoresis, and the proportion of the bacteria groups in the main bands after polymerase chain reaction-denaturing gradient gel electrophoresis was analyzed.Results The number of bands (13.57±3.37) and the diversity index of the bacteria (H' index; 2.55±0.27) in the control group were significantly higher than those in the tuberculosis group ((8.60±4.19) and 1.99±0.52)), respectively, with the statistically significant differences (t=12.55, P<0.001; t=6.75, P<0.001). A statistically significant difference (Z=-73.38, P<0.001) was found in the intra-group Dice similarity coefficient (Cs) between the control group (22.90% (16.20%, 29.30%)) and the tuberculosis group ((35.35% (23.73%, 44.98%)) (both indicate the median and quartile). Sequencing alignment showed that the main gut microbiota of the control group and the tuberculosis group were mainly Bacteroides, accounting for 73.96% (179/242) and 79.40% (158/199), respectively. The proportion of Prevotella (9.04%, 18/199) in the tuberculosis group was lower than that in the control group (24.38%, 59/242), and the difference was statistically significant (χ ²=17.82, P<0.001). The proportion of Uncultured bacterium genus in the tuberculosis group (11.56%, 23/199) was higher than that in the control group (0, 0/242), and the difference was statistically significant (χ ²=29.51, P<0.001). Conclusion The gut microbiota of healthy people and tuberculosis patients are different due to different immune functions. The occurrence of tuberculosis may result in the changes in the composition ratio of Prevotella and Uncultured bacterium.

Key words: Tuberculosis, Intestines, Bacteria, Polydeoxyribonucleotides, Denaturing gradient gel electrophoresis

Han YANG,Huan LI,Jun XI,Ai-fang LI,Ji-ru XU. Fingerprint analysis of gut microbiota by denaturing gradient gel electrophoresis in newly diagnosed tuberculosis patients[J]. Chinese Journal of Antituberculosis, 2019, 41(9): 985-992. doi: 10.3969/j.issn.1000-6621.2019.09.013

Figures/Tables 7

References 24

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肠道菌群(属/种)	对照组(242株)		结核组(199株)		χ²值	P值
肠道菌群(属/种)	菌株数	构成比(%)	菌株数	构成比(%)	χ²值	P值
Bacteroides 属	179	73.96	158	79.40	1.79	0.215
Bacteroides chinchillae	0	0.00	7	3.52	8.65	0.004
Bacteroides coprocola	12	4.96	7	3.52	0.55	0.491
Bacteroides coprophilus	8	3.31	0	0.00	6.70	0.009
Bacteroides dorei	0	0.00	27	13.57	34.98	<0.001
Bacteroides faecichinchillae	0	0.00	7	3.52	8.65	0.004
Bacteroides faecis	0	0.00	22	11.06	28.16	<0.001
Bacteroides graminisolvens	21	8.68	0	0.00	18.13	<0.001
Bacteroides helcogenes	2	0.83	0	0.00	1.65	0.504
Bacteroides koreensis	0	0.00	6	3.02	7.40	0.080
Bacteroides kribbi	0	0.00	18	9.05	22.82	<0.001
Bacteroides mediterraneensis	2	0.83	0	0.00	1.65	0.504
Bacteroides plebeius	6	2.48	0	0.00	5.00	0.035
Bacteroides sp	0	0.00	5	2.51	6.15	0.018
Bacteroides stercoris	0	0.00	10	5.03	12.44	<0.001
Bacteroides thetaiotaomicron	14	5.79	0	0.00	11.89	<0.001
Bacteroides uniformis JCM5828	51	21.07	0	0.00	47.42	<0.001
Bacteroides vulgatus	63	26.03	42	21.11	1.46	0.261
Bacteroides xylanisolvens	0	0.00	7	3.52	8.65	0.004
Prevotella属	59	24.38	18	9.04	17.82	<0.001
Romboutsia lituseburensis 属	2	0.83	0	0.00	1.65	0.504
Saccharicrinis marinus 属	2	0.83	0	0.00	1.65	0.504
Uncultured bacterium 属	0	0.00	23	11.56	29.51	<0.001
Uncultured bacterium 16sms157-2b03	0	0.00	2	1.01	2.44	0.203
Uncultured bacterium 218004-1-114	0	0.00	3	1.51	3.67	0.091
Uncultured bacterium denovo9345_591_35209	0	0.00	2	1.01	2.44	0.203
Uncultured bacterium G11_1_2c12_	0	0.00	4	2.01	4.91	0.041
Uncultured bacterium OTU561	0	0.00	2	1.01	2.44	0.203
Uncultured bacterium OTU939 16S	0	0.00	3	1.51	3.67	0.091
Uncultured bacterium YO00054E08	0	0.00	5	2.51	6.15	0.018
Uncultured Bacteroidetes bacterium	0	0.00	2	1.01	2.44	0.203