鲍曼不动杆菌生物被膜形成能力的研究

doi:10.16150/j.1671-2870.2019.05.009

摘要/Abstract

摘要：

目的：研究鲍曼不动杆菌生物被膜形成能力的影响因素,为临床防治鲍曼不动杆菌医院感染和控制其生物被膜形成提供依据。方法：收集2017年10月至12月上海交通大学医学院附属新华医院感染患者分离到的101株鲍曼不动杆菌,利用结晶紫染色法检测其生物被膜在不同温度条件下的形成能力;采用VITEK 2 Compact全自动微生物分析仪联合纸片扩散法对菌株进行药物敏感性（药敏）试验;用PCR技术检测菌株生物被膜形成相关基因（bap、abaI、bfmS、csuAB、csuA、csuC、csuD、csuE、cpaA、ompA）的携带情况。结果：在25 ℃条件下,101株鲍曼不动杆菌的生物被膜形成阳性率（93.1%）比35 ℃时（60.4%）高（P<0.05）;临床分离株对氨苄西林/舒巴坦和氨基糖苷类药物的耐药率与生物被膜形成能力间存在一定关系（P<0.05）,生物被膜形成能力为强阳性（++）的菌株对上述抗菌药物的耐药率显著低于生物被膜形成能力为阳性（+）或阴性（-）的菌株的耐药率,但是耐药率并不是随着生物被膜形成能力的增强而降低的,生物被膜形成能力为阳性（+）的菌株的耐药率反而高于形成能力为阴性（-）的菌株。鲍曼不动杆菌生物被膜形成能力的强弱与其是否携带以上10种相关基因间并无统计学相关性（P>0.05）。结论：鲍曼不动杆菌生物被膜形成能力与培养温度有关,与培养温度35 ℃相比,25 ℃条件时生物被膜形成能力高,亦与氨苄西林/舒巴坦及氨基糖苷类药物的耐药率间存在某种关联,但其与是否携带生物被膜形成相关基因（bap、abaI等）无关。

关键词: 鲍曼不动杆菌, 生物被膜, 结晶紫染色

Abstract:

Objective: To study the in vitro biofilm formation ability of Acinetobacter baumannii for providing a reference for clinical prevention and treatment of Acinetobacter baumannii infection and control of its biofilm formation. Methods: One hundred and one strains of Acinetobacter baumannii isolated from infected patients at Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine from October to December 2017 were collected. The ability of the strains to form biofilm at different temperatures was detected by crystal violet staining; the antimicrobial susceptibility test was performed by Vitek2 Compact and the disc diffusion method. The carrying status of biofilm formation-related genes (bap, abaI, bfmS, csuAB, csuA, csuC, csuD, csuE, cpaA, and ompA) was detected by PCR. Results: The positive rate of Acinetobacter baumannii biofilm formation at 25 ℃ (93.1%) was significantly higher than that at 35 ℃ (60.4%) (P<0.05). There was a correlation between the resistance rate of isolates to Ampicillin/Sulbactam or Aminoglycosides and the biofilm formation ability (P<0.05); and there was no significant correlation between the biofilm formation ability and whether it carried the abovementioned 10 related genes (P>0.05). Conclusions: The biofilm formation ability of Acinetobacter baumannii is related to the culture temperature, and also correlated in some degree with the resistance rate to Ampicillin/sulbactam and Aminoglycosides. However, the biofilm formation ability is not related to the abovementioned 10 related genes (bap, abaI, etc.).

Key words: Acinetobacter baumannii, Biofilm, Crystal violet staining

中图分类号:

R372

皇甫昱婵, 刁文晶, 俞静, 刘瑛, 沈立松. 鲍曼不动杆菌生物被膜形成能力的研究[J]. 诊断学理论与实践, 2019, 18(05): 532-537.

HUANGFU Yuchan, DIAO Wenjing, YU Jing, LIU Ying, SHEN Lisong. Study on in vitro biofilm formation ability of Acinetobacter baumannii[J]. Journal of Diagnostics Concepts & Practice, 2019, 18(05): 532-537.

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引物名称	引物序列（5'→3'）	预期产物长度(bp)
bap	F-5′-TAG GGA GGG TAC CAA TGC AG R-5′-TCA TGA TTT GAT GCT GCA GCG ATA A	405
abaI	F-5'-GTA TTT GTT GAA TAT TTG GG R-5'-GTA ATG AGT TGT TTT GCG CC	362
bfmS	F-5'-ATA TAT GCG GGG CTG GTA ATT C R-5'-ATG CAG GTG CTT TTT TAT TGG T	1 621
csuAB	F-5'-ATG AAT ATG AAA AAC ATT CA R-5'-TTA GAA ATT TAC AGT GAC TA	543
csuA	F-5'-ATG AGT AGA CCT GTT TTA AA F-5'-TTA AAA CTC AAT CGT AAT TG	579
csuC	F-5'-ATG GTG ATT TGT ATG AAC AA R-5'-TCA TGA TGG ATC CTC CAT TT	834
csuD	F-5'-TTG TCG GGT GGA TGT TTA R-5'-TTG TGG CAA TGG ATA GGT	807
csuE	F-5'-ATG CAT GTT CTC TGG ACT GAT GTT GAC R-5'-CGA CTT GTA CCG TGA CCG TAT CTT GAT AAG	977
cpaA	F-5'-CTG CTT TAG GAA AAT GGG R-5'-CGC CTT CAA TCA TTC TAA G	669
ompA	F-5'-CAA TTG TTA TCT CTG GAG R-5'-ACC TTG AGT AGA CAA ACG A	952

判断结果	35 ℃		25 ℃		P值
判断结果	菌株数	比例（%）	菌株数	比例（%）	P值
强阳性（++）	12	11.9	10	9.9	0.651
阳性（+）	49	48.5	84	83.2	<0.001
阴性（-）	40	39.6	7	6.9	<0.001

药物名称	强阳性（n=12）		阳性（n=49）		阴性（n=40）		P值^a)
药物名称	株数	耐药率（%）	株数	耐药率（%）	株数	耐药率（%）	P值^a)
哌拉西林	11	91.7	45	91.8	38	95.0	0.751
氨苄西林/舒巴坦	7	58.3	45	91.8	35	87.5	0.021
头孢哌酮/舒巴坦	4	33.3	18	36.7	15	37.5	1.000
头孢曲松	11	91.7	45	91.8	38	95.0	0.751
头孢他啶	11	91.7	45	91.8	38	95.0	0.751
美罗培南	11	91.7	44	89.8	36	90.0	1.000
亚胺培南	11	91.7	44	89.8	36	90.0	1.000
阿米卡星	6	50.0	42	85.7	32	80.0	0.034
庆大霉素	7	58.3	45	91.8	30	75.0	0.009
妥布霉素	6	50.0	42	85.7	32	80.0	0.034
环丙沙星	11	91.7	45	91.8	38	95.0	0.751
左旋氧氟沙星	5	41.7	24	49.0	18	45.0	0.874^b)
复方新诺明	7	58.3	27	55.1	27	67.5	0.505
米诺环素	0	0.0	5	10.2	6	15.0	0.388