Advanced biosensors for detection of pathogens related to livestock and poultry

Abstract

Infectious animal diseases caused by pathogenic microorganisms such as bacteria and viruses threaten the health and well-being of wildlife, livestock, and human populations, limit productivity and increase significantly economic losses to each sector. The pathogen detection is an important step for the diagnostics, successful treatment of animal infection diseases and control management in farms and field conditions. Current techniques employed to diagnose pathogens in livestock and poultry include classical plate-based methods and conventional biochemical methods as enzyme-linked immunosorbent assays (ELISA). These methods are time-consuming and frequently incapable to distinguish between low and highly pathogenic strains. Molecular techniques such as polymerase chain reaction (PCR) and real time PCR (RT-PCR) have also been proposed to be used to diagnose and identify relevant infectious disease in animals. However these DNA-based methodologies need isolated genetic materials and sophisticated instruments, being not suitable for in field analysis. Consequently, there is strong interest for developing new swift point-of-care biosensing systems for early detection of animal diseases with high sensitivity and specificity. In this review, we provide an overview of the innovative biosensing systems that can be applied for livestock pathogen detection. Different sensing strategies based on DNA receptors, glycan, aptamers and antibodies are presented. Besides devices still at development level some are validated according to standards of the World Organization for Animal Health and are commercially available. Especially, paper-based platforms proposed as an affordable, rapid and easy to perform sensing systems for implementation in field condition are included in this review.

Figure 1

Principle of biosensors. A schematic diagram of pathogen detection by a biosensor.

Figure 2

Colorimetric sensor for detection of influenza A virus. Sialic-mediated colorimetric detection of Influenza virus. Gold nanoparticles are stabilized with sialic to specifically bind HA protein on Influenza virus surface. Sialic-acid gold nanoparticles alone show the absorbance at 510 nm, while virus-bound nanoparticles absorbed at 600–610 nm. This allows label-free colorimetric readout for virus detection. Cartoon adapted from [42].

Figure 3

Lateral flow strips for detection of influenza A and B viruses. Schematic presentations of a lateral flow tests realized on a nitrocellulose strips with immobilized antibodies against influenza virus A and B. A sample containing influenza virus flow by capillarity from the sample pad to bind test and control lines. In contrast, a sample without target virus flows from the sample application pad and binds only to the control line.

Figure 4

Detection of influenza virus RNA. Available combinations of portable nucleic acid extraction, amplification and detection methods for virus detection.

Figure 5

Lateral flow kits for detection of influenza viruses. RT-RPA with lateral flow Influenza virus detection kits for influenza A, influenza B, Influenza H5 and H7 subtypes. Pictures provided from Hawk Scientific Co., Ltd and GenProNex Biomedical INC. (Flu A: influenza A, Flu B: influenza B).

Figure 6

Aptamer-based detection of influenza viruses. Schematic representations of aptamer development and virus detection. Selex procedure is applied for selection of specific aptamers. These sensing elements are immobilized on the sensor surface to bind efficiently to the viral proteins in infected samples. The recognition signal is proceeded to provide diagnostic.

Figure 7

Microarray for detection of bacterial nucleic acids. Schematic representation of microarray for mastitis bacteria detection realized on porous nitrocellulose membrane slides. Illustration adapted from Mujawar et al. [73].

Figure 8

Fluorescent detection of viral nucleic acids on magnetic beads. Schematic presentation of the FRET-based magnetic biosensor for Orbiviruses detection. (1) A nucleic acid probe labeled with florescent dye, biotin and dark quencher hybridize with the complementary probe, (2) the fluorescent dye is separate from the quencher in each PCR cycle, and starts to produce light, (3) the fluorescent dye bind to streptavidin-coated magnetic beads via biotin tag, (4) about 1000 fluorescent-labeled probes bind to a single beads giving an increased fluorescent signal. Cartoon adapted from [94].

Figure 9

Visualization and detection of Campylobacter. A Transmission electron microscopy of Campylobacter cell. Bar, 2 µm. B OLED biosensor probing of a negative control sample containing a non-Campylobacter DNA at 12.5 ng/µL, using a probe at 50 ng/µL. The spot was obtained upon deposition of 1 µL of the sample on the sensor surface. C OLED biosensor detection of Campylobacter DNA sequence at 12.5 ng/µL, using 50 ng/µL of the probe. The spot was obtained with 1 µL of the sample as previously explained [125].

Figure 10

Elements of integrated portable diagnostic laboratory. Advanced analytical technologies will offer to veterinary practitioners a suitcase containing consumables, reagents and devices to perform in field diagnostics with laboratory-grade performance.