Ligands for label-free detection of whole bacteria on biosensors: A review
Introduction
From our skin to our plates, going through all our electronic devices, such as phones and computers, microbes are everywhere around us. Although most bacteria are harmless, a few can cause various diseases ranging from minor incidence to lethal issues. Among the food-borne pathogens, Salmonella, Listeria monocytogenes and enterohemorrhagic Escherichia coli are responsible for several millions of diseases worldwide each year [1]. The World Health Organization will soon published a report estimating the Global Burden of Foodborne Diseases (http://www.who.int/foodsafety/foodborne_disease/ferg/en/ accessed on September 30, 2015). In hospital settings, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa are a considerable source of nosocomial infections. To efficiently fight these germs and reduce their impact on human health, researchers and physicians need to quickly recognize them.
Nowadays, bacteria detection still relies for the most part on classical microbiology methods including isolation and growth in selective media [2]. Although proven to be efficient, these techniques are labor intensive and time consuming due to the growth phase needed for microorganism development. It is therefore crucial to design new and innovative bacterial detection methods. To that aim, modern technics, including mass spectrometry, microarray, PCR and genomic sequencing have been intensively investigated [3]. Nevertheless, these methods generally require high technical skills, intense sample processing and rely on the presence of bacterial molecules instead of whole bacterial cells.
Over the past years, the development of biosensors has also been the focus of exhaustive researches. A biosensor is an analytical device converting a biological response into a measurable signal [4]. It is generally composed of three elements: (1) a ligand grafted on the biosensor surface which recognizes a target through specific interactions. It needs to be specific and sensitive against its target in order to induce a positive signal and prevent the interference by other substances from the sample, (2) a transducer which converts bio-recognition events arising at the surface to a physically quantifiable signal, being classified as electrical, optical, calorimetric, piezoelectric, acoustic or magnetic, (3) a detector which amplifies and analyzes the signal. Thus, biosensors convert a bio-recognition event into a physically measurable signal. The Fig. 1 gives an outline of the ligands described in this review.
Ideally, these devices need to be small, portable, easy-to-use and able to work at the point-of-care, in contrast to other diagnostic techniques. Furthermore, in order to be adapted to pathogen detection, biosensors have to give highly reproducible and rapid results. The choice of the bioreceptor is crucial for the efficiency of the biosensor, as it can influence both its sensitivity and specificity for the bacterial target. To date, a broad spectrum of bioreceptors has been used for bacterial detection. In this review, we propose an overview of the principal ligands used in biosensor systems for label-free detection of whole bacterial cells. These crucial probes are classified into three categories depending on their origin. Some ligands are natural products, or derivatives of natural products; others are engineered bio-molecules inspired from natural products whose properties are artificially improved; and eventually, some ligands are randomly synthesized biomolecules whose natural affinity for a target enables their selection from large molecular libraries. Label-free detection enables real-time measurement of the interaction and consequently requires less time and reagents than label-based methods. We will focus on whole bacteria detection, which can be directly applied on samples without the need of sample processing prior to the analysis.
Section snippets
Antibodies
Antibodies (Abs), or Immunoglobulins (Igs), are host proteins produced by the immune system of eukaryotes to neutralize and eliminate pathogens. These “Y shape” proteins are typically composed of four chains, two large heavy chains (VH) and two small light chains (VL). They possess two distinct regions, the fragment crystallizable region (Fc fragment) which interacts with and activates other immune system partners and the antigen-binding region (Fab fragment) that recognizes and binds to
Molecular Imprinted Polymers (MIP)
All the ligands listed so far in this review are described as biomolecules: proteins, peptides, sugars, nucleic acids, etc. More interestingly, these motifs are all either natural compounds (antibodies, AMP, sugars) or derived from natural products to increase their activity (engineered peptides, shortened antibodies). But since more than 30 years now, new approaches have been developed to create ligands potentially fitting any target after finger-printing engineering. This is possible by
Aptamers
Bacteria identification using DNA probes is widely used in microbiological laboratory especially linked with PCR amplification [2]. Identification relies on the binding of a DNA sequence specific of one bacterial specie with its complementary sequence found in the analyzed sample. Monitoring of the binding can be done with many transducing systems. Even if this technique is powerful, it needs a bacterial lysis step to extract the nucleic acids. Since the analysis is not carried out on the whole
Conclusions
As we have described in this review, bacterial detection systems have integrated a wide range of ligands, some of them being naturals, such as Abs and AMPs, others being engineered including aptamers, synthetic and phage peptides. Historically, antibodies have been the first choice of receptors for many biosensors and still remain extensively used. Indeed, they can be very specific to their target and often give reliable results.
Nevertheless, in some cases, the identification and development of
Acknowledgements
This work has been partially supported the “Technology for Health” program of the CEA.
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