光疗对于黄疸新生儿肠道菌群及耐药基因的影响分析

doi:10.12372/jcp.2022.21e0643

摘要/Abstract

摘要：

目的分析接受光疗的黄疸新生儿肠道菌群及其耐药基因的变化,探讨其临床意义。方法通过宏基因组测序的方法,分析黄疸新生儿接受光疗后肠道菌群及其携带耐药基因的变化。结果光疗24 h及48 h后肠道菌群丰度的α和β多样性差异无统计学意义（P>0.05）。菌种水平上,个别菌种相对丰度增多,包括梭状芽孢杆菌、粪肠球菌、嗜热链球菌（P<0.01）。新生儿肠道菌群携带大量耐药基因,共242种,主要是四环素类、β-内酰胺类、大环内酯类、磺胺类、氨基糖苷类耐药基因,同时还发现了较多大肠杆菌耐药基因。随着光疗时间延长,β-内酰胺类耐药基因（CTX-M-14）、氨基糖苷类耐药基因（aadA2）、四环素类耐药基因[tet(59)]、外排泵基因（efmA）、喹诺酮类耐药基因（QnrS2）、碳青霉烯酶基因（OXA-347）和磺胺类耐药基因（dfrA1）丰度发生显著增加;金黄色葡萄球菌调节耐药基因（mgrA）丰度在光疗后降低（P<0.05）。光疗48 h后,肠道菌群菌种水平的丰度与耐药基因呈显著正相关的有：梭状芽孢杆菌与氨基糖苷类耐药基因[APH(3')-Ia],单形拟杆菌与β-内酰胺类耐药基因（CfxA6）,阴沟肠杆菌与耐磷霉素基因（UhpT）,副血链球菌与大环内脂类耐药基因（mefA）（P<0.01）;呈显著负相关的为,粪肠球菌与氨基糖苷类耐药基因[ANT(2'')-Ia]（P<0.01）。结论光疗可导致新生儿肠道菌群部分菌种相对丰度发生变化,部分耐药基因丰度显著增加。菌群稳态被破坏,是否与光疗的不良反应及其临床意义相关需进一步研究。

关键词: 黄疸, 光疗, 肠道菌群, 耐药基因, 新生儿

Abstract:

Objective To analyze the changes of intestinal microflora and drug-resistant genes in jaundiced neonates receiving phototherapy, and to explore its clinical significance. Methods Metagenomic sequencing was used to analyze the changes of intestinal flora and drug-resistant genes in neonates with jaundice after phototherapy. Results There was no significant difference in the α- and β-diversity of the intestinal microflora abundance after 24 and 48 hours of phototherapy (P>0.05). At the species level, the relative abundance of some species increased, including Clostridium bolteae, Enterococcus faecium and Streptococcus thermophilus (P<0.01). The intestinal flora of jaundiced neonates carried a large number of drug-resistant genes, a total of 242 genes, mainly tetracycline, β-lactams, macrolides, sulfonamides and aminoglycosides, and more resistant genes of Escherichia coli were also found. With the prolongation of phototherapy time, some drug resistance genes changed significantly. The main increased resistance genes were β-lactam resistance gene (CTX-M-14), aminoglycoside resistance gene (aadA2), tetracycline resistance gene [tet(59)], efflux pump gene (efmA), quinolone resistance gene (QnrS2), carbapenemase gene (OXA-347) and sulfonamide resistance gene (dfrA1), while the abundance of mgrA in Staphylococcus aureus decreased after phototherapy, and the differences were significant (P<0.05). After 48 hours of phototherapy, the species abundance of the following intestinal microflora showed a significant positive correlation with drug resistance genes: Clostridium and aminoglycoside resistance gene [APH(3')-Ia], Bacteroides uniformis and β-lactam resistance gene (CfxA6), Enterobacter cloacae and fosfomycin resistance gene (UhpT), Streptococcus parahaemolyticus and macrolide resistance gene (mefA) (P<0.01); while the species abundance of Enterococcus faecalis was negatively correlated with aminoglycoside resistance gene [ANT(2") -Ia] (P<0.01). Conclusions Phototherapy can change the relative abundance of some bacterial species in the intestinal microflora of neonates and significantly increase the abundance of some drug-resistant genes. Whether the disruption of bacterial homeostasis is related to the adverse reactions of phototherapy and its clinical significance needs further study.

Key words: jaundice, phototherapy, intestinal microflora, drug-resistant gene, neonate

张坤, 樊赛男, 郑芳, 张佳慧, 吴志敏, 吕安平, 马亚男, 方晓慧, 张金萍. 光疗对于黄疸新生儿肠道菌群及耐药基因的影响分析[J]. 临床儿科杂志, 2022, 40(6): 436-441.

ZHANG Kun, FAN Sainan, ZHENG Fang, ZHANG Jiahui, WU Zhimin, LYU Anping, MA Yanan, FANG Xiaohui, ZHANG Jinping. The effect of phototherapy on intestinal flora and drug-resistant genes in jaundiced neonates[J]. Journal of Clinical Pediatrics, 2022, 40(6): 436-441.

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菌种	相对丰度			H值	P
菌种	光疗前	光疗24 h后	光疗48 h后	H值	P
梭状芽孢杆菌/×10^–2	10.0(5.0~81.0)	106.0(85.0~210.0)	207.7(114.2~524.6)	9.94	0.008
粪肠球菌	3.7(2.1~7.4)	4.7(2.2~9.7)	5.4 (3.3~12.0)	8.95	0.005
类杆菌	0.2(0.1~0.3)	0.2(0.1~0.4)	1.0(0.5~ 2.4)	7.26	0.034
阴沟肠杆菌	1.0(0.8~4.7)	3.0(1.0~7.2)	3.4(1.7~9.0)	8.45	0.020
巴氏链球菌	0.1(0.1~0.5)	1.7(0.8~3.3)	2.4(1.2~6.1)	7.59	0.035
嗜热链球菌/×10^–2	1.0(0.4~10.0)	3.6(1.4~19.8)	1.2(0.5~2.9)	14.59	<0.001
不动杆菌属
未分类/×10^–3	0.4(0.1~0.9)	1.3(0.7~9.6)	10.0(3.0~96.0)	6.60	0.034
丙酸棒杆菌/×10^–3	0.4(0.1~0.9)	0.1(0.1~0.8)	0.0(0.0~0.0)	4.57	0.041
直肠真杆菌/×10^–3	0.5(0~1.0)	0.9(0.2~1.2)	5.0(7.0~96.0)	9.26	0.015
痤疮丙酸杆菌/×10^–4	3.0(1.2~8.6)	0.9(0.3~3.6)	0.0(0.0~0.0)	6.60	0.037

耐药基因	光疗前	光疗24 h后	光疗48 h后	H值	P
QnrS2/×10^–3	10.0(2.0~63.3)	15.2(4.0~100.0)	20.0(30.0~26.0)	10.55	0.005
CTX-M-14	0.0(0.0~0.1)	3.3(2.6~8.7)	9.0(4.1~15.3)	9.26	0.001
dfrA1/×10^–2	50.0(5.0~90.1)	19.9(16.0~98.0)	57.7(32.2~191.0)	12.14	0.002
aadA2	0.0(0.0~0.0)	1.0(0.2~2.4)	4.7(2.6~8.3)	8.25	0.016
efmA	0.1(0.0~0.4)	0.3(0.3~1.0)	1.3(0.9~3.3)	8.25	0.013
tet(59)/×10^-2	29.9(20.1~40.1)	40.0(30.1~80.2)	85.1(50.0~120.5)	8.69	0.013
OXA-347	0.0(0.0~0.1)	0.1(0.0~0.2)	0.1(0.1~0.4)	5.45	0.031
mgrA	7.1(4.3~15.6)	5.9(2.2~9.7)	1.8(0.6~3.3)	6.09	0.048