Detection of quorum sensing molecules in Burkholderia cepacia culture supernatants with enzyme-linked immunosorbent assays |
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Authors: | Xiao Chen Katharina Buddrus-Schiemann Michael Rothballer Petra M Kr?mer and Anton Hartmann |
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Institution: | (1) Institute of Ecological Chemistry, Helmholtz Zentrum M?nchen – German Research Center for Environmental Health (GmbH), Ingolst?dter Landstrasse 1, 85764 Neuherberg, Germany;(2) Department Microbe–Plant Interactions, Helmholtz Zentrum M?nchen – German Research Center for Environmental Health (GmbH), Ingolst?dter Landstrasse 1, 85764 Neuherberg, Germany; |
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Abstract: | The Burkholderia cepacia complex (Bcc) employs a quorum sensing (QS) mechanism which is a cell density-dependent bacterial communication system to
regulate certain gene expressions. As with many other Gram-negative bacteria, Burkholderia cepacia species use (N-acyl-)homoserine lactones (AHLs or HSLs) as signalling molecules. Because of the essential role of QS in bacterial behavior,
the aim of this study was to demonstrate the applicability of our in-house-developed enzyme-linked immunosorbent assays (ELISAs)
for the detection of bacterial activities via HSLs in B. cepacia strain LA3 culture supernatants. For this purpose the previously developed monoclonal antibodies (mAbs) HSL1/2-2C10 and HSL1/2-4H5
were exploited. N-3-Oxo-decanoyl-L-homoserine lactone (3-oxo-C10-HSL) was used as main analyte throughout all experiments. With the bacterial
culture medium (named ABC medium) a matrix effect in both ELISAs was visible (slight increase in optical density, shift in
test midpoints (IC50) and working ranges). For example, ELISA with mAb HSL1/2-2C10 and enzyme tracer HSL3-HRP (HSL derivative conjugated to horseradish
peroxidase) had an IC50 of 120 μg L−1 for 3-oxo-C10-HSL in phosphate-buffered saline versus 372 μg L−1 in ABC medium. A significant increase of HSLs in B. cepacia strain LA3 culture supernatants after 12 h to 48 h of growth was observed. Although the analytical result of these immunoassays
cannot distinguish HSLs from homoserines (HSs), the appearance of these compounds can be easily followed. Hydrolysis and spiking
experiments were carried out with these biological samples. According to our knowledge, these are the first immunoassays for
the detection of quorum sensing molecules in biological culture supernatants. This study provides a cost-effective, fast,
and sensitive analytical method for detection of HSLs/HSs in biological samples without complex sample preparation and will
offer a quick idea about B. cepacia activities. The low sample amount requirement (less than 1 mL) constitutes a tremendous advantage for many analytical questions
with biological samples. |
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