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巴西西部亚马逊地区腹泻儿童中聚集性大肠杆菌的特征分析

CHARACTERIZATION OF ENTEROAGGREGATIVE ESCHERICHIA COLI AMONG DIARRHEAL CHILDRENIN WESTERN BRAZILIAN AMAZON.

作者信息

Taborda Roger Lafontaine Mesquita, Silva Luiz Antônio da, Orlandi Patricia Puccinelli, Batista Flávia Serrano, Rodrigues Renata Santos, Matos Najla Benevides

机构信息

Centro de Pesquisa em Medicina Tropical, Porto Velho, RO, Brasil.

Instituto Leônidas & Maria Deane, Fundação Oswaldo Cruz, Manaus, AM, Brasil.

出版信息

Arq Gastroenterol. 2018 Oct-Dec;55(4):390-396. doi: 10.1590/S0004-2803.201800000-84.

Abstract

BACKGROUND

Enteroaggregative Escherichia coli (EAEC) is one of the main acute and chronic diarrhea causes both in children and adults, mainly in developing countries.

OBJECTIVE

The aim of the present study is to characterize EAEC strains isolated from faecal samples and to identify genes potentially contributing to virulence, biofilm production and antimicrobial resistance in children admitted to a pediatric hospital in Porto Velho, Rondônia State.

METHODS

The total of 1,625 E. coli specimens were isolated from 591 children in the age group 6 years or younger who were hospitalized in Cosme and Damião Children Hospital in Porto Velho, between February 2010 and February 2012, with acute gastroenteritis. Colonies suggestive of E. coli were subjected to polymerase chain reaction testing in order to identify the virulence factors. The in vitro adhesion assays using HEp-2 adherence were tests. Biofilm detection through spectrophotometry and antimicrobial susceptibility tests were conducted in the disk diffusion method.

RESULTS

The mentioned study examined 591 stool samples from children with diarrhea. Diarrheogenic E. coli was found in 27.4% (162/591) of the children. EAEC was the diarreagenic E. coli most frequently associated with diarrhea 52.4% (85/162), which was followed by enteropathogenic E. coli 43.8% (71/162), enterotoxigenic E. coli 2.4% (4/162), and enterohemorrhagic E. coli 1.2% (2/162). The aggR gene was detected in 63.5% (54/85) of EAEC isolates; moreover, statistically significant correlation was observed among typical EAEC (aggR) and aatA (P<0.0001), irp2 (P=0.0357) and shf (P=0.0328). It was recorded that 69% (59/85) of the 85 analyzed EAEC strains were biofilm producers; 73% (43/59) of the biofilm producers carried the aggR gene versus 42.3% (11/26) of non-producers (P=0.0135). In addition, there was association between the aatA gene and biofilm production; 61% (36/59) of the samples presented producer strains, versus 19.2% (5/26) of non-producers (P<0.0004). Antibiotic sensitivity test evidenced that most EAEC were ampicillin 70.6% (60/85), sulfamethoxazole 60% (51/85), tetracycline 44.7% (38/85) and cefotaxime 22.4% (19/85) resistant.

CONCLUSION

As far as it is known, the present study is pioneer in Northern Brazil to investigate EAEC virulence factors and to show the antimicrobial susceptibility of EAEC strains isolated from children with diarrhea.

摘要

背景

肠聚集性大肠杆菌(EAEC)是儿童和成人急性及慢性腹泻的主要病因之一,主要见于发展中国家。

目的

本研究旨在对从粪便样本中分离出的EAEC菌株进行特征分析,并鉴定可能与罗赖马州波多韦柳市一家儿科医院收治儿童的毒力、生物膜形成及抗菌药物耐药性相关的基因。

方法

2010年2月至2012年2月期间,从在波多韦柳市科斯梅和达米亚昂儿童医院住院的591名6岁及以下患急性肠胃炎的儿童中分离出1625株大肠杆菌样本。对疑似大肠杆菌的菌落进行聚合酶链反应检测以鉴定毒力因子。采用HEp-2黏附法进行体外黏附试验。通过分光光度法检测生物膜,并采用纸片扩散法进行抗菌药物敏感性试验。

结果

上述研究检测了591份腹泻儿童的粪便样本。在27.4%(162/591)的儿童中发现了致泻性大肠杆菌。EAEC是最常与腹泻相关的致泻性大肠杆菌,占52.4%(85/162),其次是肠致病性大肠杆菌,占43.8%(71/162),产肠毒素大肠杆菌占2.4%(4/162),肠出血性大肠杆菌占1.2%(2/162)。在63.5%(54/85)的EAEC分离株中检测到aggR基因;此外,在典型EAEC(aggR)与aatA(P<0.0001)、irp2(P=0.0357)和shf(P=0.0328)之间观察到统计学显著相关性。记录显示,在分析的85株EAEC菌株中,69%(59/85)是生物膜产生菌;生物膜产生菌中有73%(43/59)携带aggR基因,而非产生菌中这一比例为42.3%(11/26)(P=0.0135)。此外,aatA基因与生物膜形成之间存在关联;样本中有61%(36/59)呈现产生菌菌株,而非产生菌中这一比例为19.2%(5/26)(P<0.0004)。抗生素敏感性试验证明,大多数EAEC对氨苄西林耐药率为70.6%(60/85),对磺胺甲恶唑耐药率为60%(51/85),对四环素耐药率为44.7%(38/85),对头孢噻肟耐药率为22.4%(19/85)。

结论

据了解,本研究是巴西北部首个调查EAEC毒力因子并展示从腹泻儿童中分离出的EAEC菌株抗菌药物敏感性的研究。

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