Quantitative Detection of Mastitis Factor IL-6 in Dairy Cow Using the SERS Improved Immunofiltration Assay

Abstract

Interleukin-6 (IL-6) is generally used as a biomarker for the evaluation of inflammatory infection in humans and animals. However, there is no approach for the on-site and rapid detection of IL-6 for the monitoring of mastitis in dairy farm scenarios. A rapid and highly sensitive surface enhanced Raman scattering (SERS) immunofiltration assay (IFA) for IL-6 detection was developed in the present study. In this assay, a high sensitivity gold core silver shell SERS nanotag with Raman molecule 4-mercaptobenzoic acid (4-MBA) embedded into the gap was fabricated for labelling. Through the immuno-specific combination of the antigen and antibody, antibody conjugated SERS nanotags were captured on the test zone, which facilitated the SERS measurement. The quantitation of IL-6 was performed by the readout Raman signal in the test region. The results showed that the detection limit (LOD) of IL-6 in milk was 0.35 pg mL−1, which was far below the threshold value of 254.32 pg mL−1. The recovery of the spiking experiment was 87.0-102.7%, with coefficients of variation below 9.0% demonstrating high assay accuracy and precision. We believe the immunosensor developed in the current study could be a promising tool for the rapid assessment of mastitis by detecting milk IL-6 in dairy cows. Moreover, this versatile immunosensor could also be applied for the detection of a wide range of analytes in dairy cow healthy monitoring.

A Rapid and Inexpensive PCR Test for Mastitis Diagnosis Based on NGS Data

“…The literature suggests various immunoassay tests for early inflammation screening utilizing biomarkers like milk amyloid A [40], cathelicidins [41], myeloperoxidase enzyme of milk neutrophils [42], IL-6 [43] as well as lateral flow assays for diagnosing mastitis [44]. While the abovementioned methods offer relatively low costs and high precision in com-parison to standard diagnostic methods, we cannot obtain results indicating the bacteria species, which is crucial for the implementation of appropriate antibiotic therapy, thus it can be used as a screening test, but still, microbiological and genetic tests should be conducted.…”

Diagnosis of Subclinical Staphylococcal Aureus Bovine Mastitis Using Nanotechnology-Based Techniques

“…Interleukin-6 (IL-6) is employed as a biomarker to assess inflammatory infections in both humans and animals and the immunofiltration assay (IFA) has been extensively utilized in the field of inflammatory biomarker identification to facilitate the sensitive and quantitative measurement of IL-6 in milk [55]. a gold-silver core-shell surface-enhanced Raman scattering (SERS) nanotag with 4-mercaptobenzoic acid 4-MBA was used for labeling in the immune filtration assay (IFA) By using an immuno-specific combination of antigen and antibody [55]. Surfaceenhanced Raman scattering (SERS) has gained more interest due to a quick response time, no interference from an aqueous solution, minimal sample pretreatment needed, high accuracy, and no sample degradation [56].…”

Insights into the Prognostic Role of Serum Interleukin-6 and Hematobiochemical Alterations in Cattle during Recent Outbreaks of Lumpy Skin Disease in Lodhran District, Pakistan

“…Interleukin-6 (IL-6), known as a multifunctional cytokine, is a glycoprotein generated and secreted by lymphocytes that activates macrophages and enhances proinflammatory response [ 14 ]. The prognostic aspect of IL-6 has been investigated in cattle in several studies [ 15 , 16 , 17 ]. However, limited information is available regarding serum IL-6 levels in the case of LSD.…”