Abstract

BackgroundEarly diagnosis and classification of infections increase the cure rate while decreasing complications, which is significant for severe infections, especially for war surgery. However, traditional methods rely on laborious operations and bulky devices. On the other hand, point-of-care (POC) methods suffer from limited robustness and accuracy. Therefore, it is of urgent demand to develop POC devices for rapid and accurate diagnosis of infections to fulfill on-site militarized requirements.MethodsWe developed a wave-shaped microfluidic chip (WMC) assisted multiplexed detection platform (WMC-MDP). WMC-MDP reduces detection time and improves repeatability through premixing of the samples and reaction of the reagents. We further combined the detection platform with the streptavidin–biotin (SA-B) amplified system to enhance the sensitivity while using chemiluminescence (CL) intensity as signal readout. We realized simultaneous detection of C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) on the detection platform and evaluated the sensitivity, linear range, selectivity, and repeatability. Finally, we finished detecting 15 samples from volunteers and compared the results with commercial ELISA kits.ResultsDetection of CRP, PCT, and IL-6 exhibited good linear relationships between CL intensities and concentrations in the range of 1.25–40 μg/ml, 0.4–12.8 ng/ml, and 50–1600 pg/ml, respectively. The limit of detection of CRP, PCT, and IL-6 were 0.54 μg/ml, 0.11 ng/ml, and 16.25 pg/ml, respectively. WMC-MDP is capable of good adequate selectivity and repeatability. The whole detection procedure takes only 22 min that meets the requirements of a POC device. Results of 15 samples from volunteers were consistent with the results detected by commercial ELISA kits.ConclusionsWMC-MDP allows simultaneous, rapid, and sensitive detection of CRP, PCT, and IL-6 with satisfactory selectivity and repeatability, requiring minimal manipulation. However, WMC-MDP takes advantage of being a microfluidic device showing the coefficients of variation less than 10% enabling WMC-MDP to be a type of point-of-care testing (POCT). Therefore, WMC-MDP provides a promising alternative to POCT of multiple biomarkers. We believe the practical application of WMC-MDP in militarized fields will revolutionize infection diagnosis for soldiers.

Highlights

  • Diagnosis and classification of infections increase the cure rate while decreasing complications, which is significant for severe infections, especially for war surgery

  • High levels of C-reactive protein (CRP) are related to bacterial infections and some viral infections like Hemagglutinin 1 Neuraminidase 1 (H1N1) and Corona Virus Disease 2019 (COVID-19) [12,13,14]

  • Detection antibodies of IL-6 conjugated with Biotin (B-IL-6-Detection antibodies of PCT (Ab2)) and horseradish peroxidase (HRP) conjugated with SA (SA-HRP) were obtained from Thermo Fisher Scientific (USA)

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Summary

Introduction

Diagnosis and classification of infections increase the cure rate while decreasing complications, which is significant for severe infections, especially for war surgery. Multiplex detection of C-reactive protein (CRP) [7], procalcitonin (PCT) [8], and interleukin-6 (IL-6) [9] is a reliable method for accurately diagnosing infections and sepsis [10]. Coronary heart disease [15, 16], cancer [17, 18], and other diseases of such kind have high concentrations of CRP For this reason, diagnoses of infections relying on the detection of CRP are not accurate. Multiplexed detections of CRP, PCT, and IL-6 can cover the whole period of infection. It effectively assesses the classification and severity of infections [21]. It is imperative to develop a multiplexed detection technique for infectious biomarkers in the fields of militarized point-of-care testing (POCT) [22,23,24]

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