In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environmentsReport as inadecuate




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BMC Microbiology

, 16:209

Microbe-host interactions and microbial pathogenicity

Abstract

BackgroundThere is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the host’s defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms.

The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms.

MethodsLymphocytes were separated from whole blood of patients n = 10 and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system SPR and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies.

ResultsThe produced antibodies recognized bacteria with high sensitivity SPR. The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy.

ConclusionIn the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.

KeywordsEnterococcus Biofilms Antibodies Detection Electron microscopy Lymphocytes Surface plasmon resonance AbbreviationsE.colEscherichia coli

E.fecalisEnterococcus fecalis

EDSN-ethyl-N-3 diethylaminopropyl carboiimide

ESBLExtended spectrum beta-lactamase

NaclNatrium chloride

NHSN-hydroxysuccinimide

NPVNegative predictive value

P. aeruginosaPseudomonas aeruginosa

P. gingivalisPorphyromonas gingivalis

PBSPhosphate buffer solution

PPVPositive predictive value

RUResponse unit

S. aureusStaphylococcus aureus

S. epidermidisStaphylococcus epidermidis

SPRSurface plasmon resonance

Electronic supplementary materialThe online version of this article doi:10.1186-s12866-016-0821-5 contains supplementary material, which is available to authorized users.

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Author: Chongcong Wu - Sravya Nakka - Sepahdar Mansouri - Torbjörn Bengtsson - Tayeb Nayeri - Fariba Nayeri

Source: https://link.springer.com/







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