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印度北部分离的多重耐药性尿路致病性肠球菌的生物膜合成及其他毒力因子。

Biofilm synthesis and other virulence factors in multidrug-resistant uropathogenic enterococci isolated in Northern India.

机构信息

Department of Microbiology, Hamdard Institute of Medical Sciences and Research, Jamia Hamdard, New Delhi, India.

Department of Medicine, Hamdard Institute of Medical Sciences and Research, Jamia Hamdard, New Delhi, India.

出版信息

Indian J Med Microbiol. 2020 Apr-Jun;38(2):200-209. doi: 10.4103/ijmm.IJMM_19_355.

Abstract

PURPOSE

Enterococci express high degree of resistance towards wide range of antibiotics. Production of biofilm and many virulence factors along with drug resistance makes it difficult to eradicate the infection from urinary tract. The present study detected the expression of such factors including biofilm production by multidrug-resistant (MDR) enterococci.

MATERIALS AND METHODS

Drug susceptibility of 103 uropathogenic enterococci was performed followed by estimation of minimum inhibitory concentration of high-level gentamicin and vancomycin by microbroth dilution method. Vancomycin-resistant genes were detected by multiplex polymerase chain reaction. Production of virulence factors such as haemagglutination, caseinase, lipase, gelatinase, haemolysin and β-lactamase was detected by phenotypic methods in MDR strains. Biofilm production was detected by calcofluor-white fluorescence staining and semi-quantitative adherence assay.

RESULTS

45% and 18.4% of the isolates were high-level gentamicin-resistant and vancomycin-resistant enterococci (VRE), respectively. vanA gene was detected in 14 and vanB gene in 5 strains. Biofilm, caseinase and gelatinase were the most expressed virulence factor. Expression of caseinase, gelatinase and lipase was significantly higher in Enterococcus faecalis (P < 0.05). Expression of haemagglutination, gelatinase and haemolysin among the vancomycin-resistant isolates was significantly higher (P < 0.05).

CONCLUSION

VanA and vanB are the prevalent genotypes responsible for vancomycin resistance. The high prevalence of MDR enterococcal strains producing biofilm and virulence determinants raises concern. asa1, hyl, esp, gelE, cyl and other genes are known to express these factors and contribute to biofilm formation. Most uropathogenic enterococci expressed biofilm at moderate level and can be detected effectively by calcofluor-white staining. No correlation was noted between vancomycin resistance and biofilm production.

摘要

目的

肠球菌对多种抗生素表现出高度耐药性。生物膜的产生和许多毒力因子以及耐药性使得从尿路感染中根除感染变得困难。本研究检测了包括多药耐药(MDR)肠球菌生物膜产生在内的这些因子的表达。

材料和方法

对 103 株尿路致病性肠球菌进行了药物敏感性检测,然后采用微量肉汤稀释法测定高水平庆大霉素和万古霉素的最小抑菌浓度。采用多重聚合酶链反应检测万古霉素耐药基因。采用表型方法检测 MDR 株产毒力因子,如血凝、胞酶、脂肪酶、明胶酶、溶菌素和β-内酰胺酶。采用钙荧光白荧光染色和半定量粘附试验检测生物膜的产生。

结果

45%和 18.4%的分离株分别为高水平庆大霉素耐药和万古霉素耐药肠球菌(VRE)。14 株菌中检测到 vanA 基因,5 株菌中检测到 vanB 基因。生物膜、胞酶和明胶酶是表达最丰富的毒力因子。粪肠球菌中胞酶、明胶酶和脂肪酶的表达显著更高(P<0.05)。万古霉素耐药分离株中血凝、明胶酶和溶菌素的表达显著更高(P<0.05)。

结论

vanA 和 vanB 是导致万古霉素耐药的主要基因型。高流行率的产生物膜和毒力决定因子的 MDR 肠球菌菌株引起关注。asa1、hyl、esp、gelE、cyl 和其他基因已知表达这些因子并有助于生物膜形成。大多数尿路致病性肠球菌以中等水平表达生物膜,并用钙荧光白染色可有效检测。万古霉素耐药与生物膜产生之间无相关性。

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