Analytical Performance of a FRET-Based Point-of-Care Immunoassay for the Quantitation of C-Reactive Protein in Serum and Finger Prick Capillary Whole Blood

<div>Time-resolved fluorescence resonance energy transfer (FRET) provides a useful technology for immunoassay measurement of proteins, This has been used to develop an immunoassay for C-reactive protein (Procise CRP<a>™</a>) that provides point-of-care measurement of CRP in less than 4 minutes using 20 µL of either whole blood or serum. Analytical studies of the assay on the ProciseDx™ analyzer were performed to characterize its measurement capabilities. Sensitivity, specificity, linearity, and precision - including reproducibility of finger prick blood collection and testing, suitable for routine clinical use in a point-of-care setting were demonstrated for the CRP assay. It also showed excellent analytical agreement with a current commercial CRP test. These results indicate the Procise CRP assay is useful for obtaining fast and accurate CRP quantitation in a point of care setting that can aid in the immediate assessment of patients’ inflammatory status. </div><div> </div>

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Development of a point-of-care assay system for high-sensitivity C-reactive protein in whole blood

Clinica Chimica Acta ◽

10.1016/s0009-8981(03)00113-x ◽

2003 ◽

Vol 332 (1-2) ◽

pp. 51-59 ◽

Cited By ~ 67

Author(s):

Jae Soon Ahn ◽

Sunga Choi ◽

Sang Ho Jang ◽

Hyuk Jae Chang ◽

Jae Hoon Kim ◽

...

Keyword(s):

Whole Blood ◽

Point Of Care ◽

High Sensitivity ◽

Assay System ◽

C Reactive Protein ◽

Reactive Protein ◽

Point Of Care Assay

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Rapid C-reactive protein determination in whole blood with a White Light Reflectance Spectroscopy label-free immunosensor for Point-of-Care applications

Sensors and Actuators B Chemical ◽

10.1016/j.snb.2018.01.008 ◽

2018 ◽

Vol 260 ◽

pp. 282-288 ◽

Cited By ~ 8

Author(s):

Georgios Koukouvinos ◽

Dimitrios Goustouridis ◽

Konstantinos Misiakos ◽

Sotirios Kakabakos ◽

Ioannis Raptis ◽

...

Keyword(s):

White Light ◽

Whole Blood ◽

Point Of Care ◽

Reflectance Spectroscopy ◽

Label Free ◽

C Reactive Protein ◽

Protein Determination ◽

Reactive Protein ◽

Light Reflectance

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Commutability of a whole-blood external quality assessment material for point-of-care C-reactive protein, glucose, and hemoglobin testing

Clinica Chimica Acta ◽

10.1016/j.cca.2019.03.1365 ◽

2019 ◽

Vol 493 ◽

pp. S647

Author(s):

T. Bukve

Keyword(s):

Quality Assessment ◽

Whole Blood ◽

External Quality Assessment ◽

Point Of Care ◽

C Reactive Protein ◽

External Quality ◽

Reactive Protein

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Ultrarapid, Ultrasensitive One-Step Kinetic Immunoassay for C-Reactive Protein (CRP) in Whole Blood Samples: Measurement of the Entire CRP Concentration Range with a Single Sample Dilution

Clinical Chemistry ◽

10.1093/clinchem/48.2.269 ◽

2002 ◽

Vol 48 (2) ◽

pp. 269-277 ◽

Cited By ~ 12

Author(s):

Piia Tarkkinen ◽

Tom Palenius ◽

Timo Lövgren

Keyword(s):

Whole Blood ◽

Solid Phase ◽

Point Of Care ◽

High Sensitivity ◽

Clinical Samples ◽

Cardiac Complications ◽

Fluorescence Signal ◽

C Reactive Protein ◽

Reactive Protein ◽

One Step

Abstract Background: Recently, measurement of very low concentrations of C-reactive protein (CRP) has gained popularity as a potential new means for predicting the risk of future cardiac complications. In this study, we demonstrate the feasibility of a kinetic, one-step microparticle assay for quantitative determination of extremely low and high CRP concentrations in the limited timeframe typical for point-of-care testing. Methods: A noncompetitive, kinetic CRP immunoassay was developed that uses individual, porous microparticles as the solid phase. The microparticles were covalently coated with a monoclonal capture antibody, and the monoclonal detection antibody was labeled with europium. The one-step binding reaction was stopped by washing after 2 min of incubation, and the fluorescence signal of individual particles was measured. Results: The analytical detection limit (mean of zero calibrator + 3 SD) was 0.00016 mg/L CRP. Clinical samples were diluted 400-fold before assay to cover the CRP concentration range of 0.064–1200 mg/L. The assay correlated well with the Dade Behring N High Sensitivity CRP assay (for 0–10 mg/L, r = 0.969, Sy|x = 0.68, n = 54; for 0–350 mg/L, r = 0.969, Sy|x = 11.7, n = 100). The within- and between-run CVs based on calculated concentrations were, respectively, 9–16% and 14% at 0.11 mg/L, 4.5–12% and 8.2% at 4.2 mg/L, and 3.5–6.3% and 4.4% at 105 mg/L, with a CV <15% at 0.2 mg/L and above. Conclusions: Use of the kinetic microparticle approach combined with time-resolved fluorometry allows ultrasensitive quantification of CRP in whole blood in 2 min with a linear assay range spanning more than four orders of magnitude.

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Therapeutic Drug Monitoring: Performance of a FRET-Based Point-Of-Care Immunoassay for the Quantitation of Infliximab and Adalimumab in Blood

10.26434/chemrxiv.13277321.v1 ◽

2020 ◽

Author(s):

Edgar Ong ◽

Ruo Huang ◽

Richard Kirkland ◽

Stefan Westin ◽

Jared Salbato ◽

...

Keyword(s):

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Whole Blood ◽

Point Of Care ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Drug Level ◽

Analytical Performance ◽

Quantitative Detection ◽

Therapeutic Drug

Two fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassays (Procise IFX™ and Procise ADL™) were developed for the quantitative detection of infliximab (IFX), adalimumab (ADL), and their respective biosimilars for use in therapeutic drug monitoring (TDM) using 20 µL of finger prick whole blood at the point-of-care or whole blood/serum in a central lab. Studies were performed to characterize analytical performance of the Procise IFX and the Procise ADL assays on the ProciseDx™ analyzer. The Procise IFX and Procise ADL assays both showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use as well as excellent correlation to current commercial ELISA IFX and ADL measurement methods. Results indicated that the Procise IFX and Procise ADL assays are sensitive, specific, and precise yielding results in less than 5 minutes from either whole blood or serum. This indicates the Procise IFX and Procise ADL assays are useful for obtaining fast and accurate IFX or ADL quantitation, thus avoiding delays inherent to current methods and enabling immediate drug level dosing decisions to be made during a single patient visit.

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P544 Point-of-care finger prick test for C-reactive protein, infliximab and adalimumab serum concentrations

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjab076.665 ◽

2021 ◽

Vol 15 (Supplement_1) ◽

pp. S514-S515

Author(s):

A Volkers ◽

M Löwenberg ◽

K Bray ◽

B Bahur ◽

M Braad ◽

...

Keyword(s):

Point Of Care ◽

Serum Concentrations ◽

C Reactive Protein ◽

Prick Test ◽

Reactive Protein ◽

Drug Concentrations ◽

Deming Regression ◽

Finger Prick ◽

Point Of Care Tests ◽

Serum Crp

Abstract Background Point-of-Care tests (POCTs) allow instant measurement of inflammatory markers and/or drug concentrations. However, currently available POCTs for infliximab (IFX) and adalimumab (ADL) serum concentrations are time consuming. Recently, a new POCT device (ProciseDx, San Diego, CA, USA) was developed which conveniently measures C-reactive protein (CRP), IFX and ADL capillary concentrations within minutes. We aimed to validate this device by comparing POCT results with conventional laboratory tests for serum CRP, IFX and ADL in patients with inflammatory bowel disease (IBD). Methods IBD patients requiring routine measurement of serum CRP, IFX or ADL were invited to participate. Along with serum collection, 20μl of capillary blood was obtained via finger prick and dispensed in a cartridge with a buffer, and placed in the POCT device providing results within two to four minutes. Forty patients were needed to validate the CRP POCT, as this assay was already previously validated in a different population. For the IFX and ADL assays, 120 patients using IFX or ADL were required to validate the POCT. Agreement between the laboratory serum assay and POCT was visualized on a scatter diagram and a Bland-Altman plot. Deming regression was calculated to demonstrate agreement. In addition, Pearson’s correlation coefficient was calculated. Results Until now, 41 patients have been enrolled for the CRP assay, 120 patients for the IFX and 46 patients for the ADL assay. Two ADL patients were sampled twice (n=48). Significant correlations were found for CRP, IFX and ADL (r=0.98, r=0.88 and r=0.86 respectively, Fig. 1). Deming regression analysis of the CRP assay resulted in a slope of 0.71 (95% CI 0.49 to 0.93) and 1.5 (95% CI -0.44 to 3.50) for the intercept. For the IFX assay, the slope was 1.1 (95% CI 0.83 to 1.3) and the intercept was 1.4 (95% CI -0.47 to 3.4). For the ADL assay, the slope was 0.97 (95% CI 0.64 to 1.3) and the intercept was 2.3 (95% CI -0.64 to 5.2). Conclusion A novel POCT using a finger prick approach provides a rapid, user friendly and reliable measurement of CRP, IFX and ADL concentrations within minutes. The capillary CRP was slightly lower than the venous serum CRP, which was consistently observed and considered clinically irrelevant in this cohort.

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P216 Comparative Assessment C-reactive Protein Between a Point-of-Care Testing and Current Standard of Care (Immunonephelometric testing)

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjab076.342 ◽

2021 ◽

Vol 15 (Supplement_1) ◽

pp. S272-S273

Author(s):

G Lorenzon ◽

I Marsilio ◽

D Maniero ◽

A Rigo ◽

B Barberio ◽

...

Keyword(s):

Inflammatory Disease ◽

Point Of Care ◽

Standard Of Care ◽

Turnaround Time ◽

Blood Collection ◽

Standard Laboratory ◽

Point Of Care Testing ◽

C Reactive Protein ◽

Serum Samples ◽

Reactive Protein

Abstract Background C-reactive protein (CRP) is widely used as a biomarker of inflammatory disease activity in hospitalized and non-hospitalized patients. In particular, CRP is commonly used in patients suspected to have an inflammatory bowel disease (IBD) or with a confirmed diagnosis of IBD diagnosis in order to drive the diagnostic approach, to monitor disease activity and to guide therapeutic adjustments. However, standard laboratory CRP testing (Immunonephelometric assays) present some drawbacks, including a turnaround time of 1–2 hours, and the need of specialized equipment, offices and laboratory personnel. Because of that, point-of care testing (POCT) was recently developed in order to provide results within 2 minutes from blood collection, enabling a rapid response to clinical condition. Aim To determine the degree of analytical correlation between a recently developed POCT (ProciseDx) using capillary whole blood and the comparative Immunonephelometric assay using serum samples. Methods From October to November 2020, consecutive patients hospitalized at Gastroenterology Unit, Padua University Hospital, aged > 18 years and with clinical evidence of active inflammatory disease or infection, who underwent to a standard of care CRP test (Dimension Vista – Siemens Healthineers) were included in the study (range 2.9–340 g/L). Within 1 hour from blood collection, in each patient, CRP quantitation from capillary whole blood collected by finger stick was performed using the ProciseDx CRP assay, with reportable range between 3.6–100 g/L. A Deming regression test was used to identify the correlation between the two methods. Results Eighty-three patients were enrolled (62.5% males with mean age ± SD: 60±18). The most common indications for hospitalisation were liver disease (34.9%), pancreatic disturbance (27.7%) and suspicious or recurrence of IBD (16.7%). ProciseDx POCT with finger prick samples required a turnaround time of 2±0.2 minutes, whereas serum samples analyzed in clinical laboratory with the reference method required a turnaround time of about 180±15 minutes (p<0.001). Overall, the correlation between the two tests was high (R squared of 0.899 (95% CI 0.916–0.968)). In particular, the correlation between the methods was even higher with CRP values between 0–100 g/L with R squared of 0.961 (95% CI 0.958–0.986). Conclusion The ProciseDx POCT allows a more rapid and comparable accuracy of CRP assessment in hospitalized patients as compared to the standard laboratory measurement. Moreover, the ProciseDx POCT does not require specialised personnel to be performed. The use of ProciseDx POCT may improve and accelerate the decision-making approach, further reducing the resources required for CRP assessment.

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Clinical Value of Whole Blood Procalcitonin Using Point of Care Testing, Quick Sequential Organ Failure Assessment Score, C-Reactive Protein and Lactate in Emergency Department Patients with Suspected Infection

Journal of Clinical Medicine ◽

10.3390/jcm8060833 ◽

2019 ◽

Vol 8 (6) ◽

pp. 833 ◽

Cited By ~ 1

Author(s):

Bo-Sun Shim ◽

Young-Hoon Yoon ◽

Jung-Youn Kim ◽

Young-Duck Cho ◽

Sung-Jun Park ◽

...

Keyword(s):

Emergency Department ◽

Whole Blood ◽

Predictive Value ◽

Point Of Care ◽

Point Of Care Testing ◽

C Reactive Protein ◽

Suspected Infection ◽

Reactive Protein ◽

Failure Assessment ◽

Emergency Department Patients

We investigated the clinical value of whole blood procalcitonin using point of care testing, quick sequential organ failure assessment score, C-reactive protein and lactate in emergency department patients with suspected infection and assessed the accuracy of the whole blood procalcitonin test by point-of-care testing. Participants were randomly selected from emergency department patients who complained of a febrile sense, had suspected infection and underwent serum procalcitonin testing. Whole blood procalcitonin levels by point-of-care testing were compared with serum procalcitonin test results from the laboratory. Participants were divided into two groups—those with bacteremia and those without bacteremia. Sensitivity, specificity, positive predictive value, negative predictive value of procalcitonin, lactate and Quick Sepsis-related Organ Failure Assessment scores were investigated in each group. Area under receiving operating curve of C-reactive protein, lactate and procalcitonin for predicting bacteremia and 28-day mortality were also evaluated. Whole blood procalcitonin had an excellent correlation with serum procalcitonin. The negative predictive value of procalcitonin and lactate was over 90%. Area under receiving operating curve results proved whole blood procalcitonin to be fair in predicting bacteremia or 28-day mortality. In the emergency department, point-of-care testing of whole blood procalcitonin is as accurate as laboratory testing. Moreover, procalcitonin is a complementing test together with lactate for predicting 28-days mortality and bacteremia for patients with suspected infection.

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Therapeutic Drug Monitoring: Performance of a FRET-Based Point-Of-Care Immunoassay for the Quantitation of Infliximab and Adalimumab in Blood

10.26434/chemrxiv.13277321 ◽

2020 ◽

Author(s):

Edgar Ong ◽

Ruo Huang ◽

Richard Kirkland ◽

Stefan Westin ◽

Jared Salbato ◽

...

Keyword(s):

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Whole Blood ◽

Point Of Care ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Drug Level ◽

Analytical Performance ◽

Quantitative Detection ◽

Therapeutic Drug

Two fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassays (Procise IFX™ and Procise ADL™) were developed for the quantitative detection of infliximab (IFX), adalimumab (ADL), and their respective biosimilars for use in therapeutic drug monitoring (TDM) using 20 µL of finger prick whole blood at the point-of-care or whole blood/serum in a central lab. Studies were performed to characterize analytical performance of the Procise IFX and the Procise ADL assays on the ProciseDx™ analyzer. The Procise IFX and Procise ADL assays both showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use as well as excellent correlation to current commercial ELISA IFX and ADL measurement methods. Results indicated that the Procise IFX and Procise ADL assays are sensitive, specific, and precise yielding results in less than 5 minutes from either whole blood or serum. This indicates the Procise IFX and Procise ADL assays are useful for obtaining fast and accurate IFX or ADL quantitation, thus avoiding delays inherent to current methods and enabling immediate drug level dosing decisions to be made during a single patient visit.

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