Development and Evaluation of Europium-Based Quantitative Lateral Flow Immunoassay for the Chronic Kidney Disease Marker Cystatin-C

Author(s):  
Satheesh Natarajan ◽  
Ebru Saatçi ◽  
Jayaraj Joseph
2021 ◽  
Author(s):  
SATHEESH NATARAJAN ◽  
Ebru saatci

Abstract This study aimed to establish a Europium label time-resolved fluorescence immunoassay (TRFIA) to detect the chronic kidney disease (CKD) biomarker Cystatin-C. Some Cystatin-c immunoassays are sensitive, accurate, and available for clinical application, but they are expensive and time-consuming procedures. Also, conventional organic dye-based fluorescence lateral flow assay showed more background fluorescence interference and low analytical sensitivity. So this Europium-based sandwich immunoassay was developed to detect the concentration of cystatin-c in a urine sample with captured anti-cystatin-c antibodies immobilized on nitrocellulose membrane and then bonded with detection anti-cystatin-c labelled with CM-EU, followed by fluorescence measurement using time-resolved fluorometry in 15 minutes. The performance of this TRFIA was evaluated using the clinical urine serum and compared with the ELISA assays. The linear calibration range was 0.015-32 µg/ml, and the limit of detection (LOD) quantified was 0.0001µg/ml. This current work has improved the LOD of our previous work from 0.013µg/ml to 0.001µg/ml. These results indicated that the CM-EU nanoparticle-based LFIA is rapid, more sensitive, reliable, and reproducible for point-of-care testing of Cys-C concentrations in urine


2009 ◽  
Vol 112 (3) ◽  
pp. c164-c170 ◽  
Author(s):  
Néstor Fontseré ◽  
Vicens Esteve ◽  
Ana Saurina ◽  
Mónica Pou ◽  
Nuria Barba ◽  
...  

2010 ◽  
Vol 140 ◽  
pp. S33
Author(s):  
Serkan Ordu ◽  
Ismail Erden ◽  
Recai Alemdar ◽  
Mesut Aydin ◽  
Onur Caglar ◽  
...  

2018 ◽  
Vol 35 (1-2) ◽  
pp. 43-54
Author(s):  
Alyaa A. Kotby ◽  
Waleed M. El Guindy ◽  
Mohamed S. El Farsy ◽  
Nanies M. S. Soliman ◽  
Menat Allah A. Shaaban ◽  
...  

2021 ◽  
pp. 189-190
Author(s):  
G.G. Kaushik ◽  
Shubham Maheshwari ◽  
Ankita Sharma

Introduction: Serum lipocalin 2 serve as a marker for kidney function. Lipocalin 2 is found in both CKD and kidney injury and it rises in acute kidney injury (AKI) and in patients have faster decline in kidney function. Aims And Objectives: To nd out correlation and assess of serum Neutrophil gelatinase-associated lipocalin 2 (NGAL 2) in patients with stages 2 to 4 of Chronic Kidney disease. The aim of the study was NGAL could represent a novel, sensitive marker of kidney function in adult patients with CKD. Material And Methods: Study involved 120 patients divided in Case group (60 patients) attended medical/ urology OPD or admitted in medical/urology ward of CKD2 – CKD4 while control group – age and sex matched healthy individuals/ stage I CKD patients was taken as control. The plasma/ serum were used for serum urea, creatinine, Cystatin C and lipocalin 2 under all aseptic precaution on receiving consent. Result:The patients of CKD included in study were having glomerulonephritis (46.7%), pyelonephritis (21.7%), diabetic kidney disease (13.3%), polycystic kidney disease (1.7%) and other causes (16.7%). CKD patients demonstrated elevated serum NGAL 159.14 ± 48.73 ng/ml, together with a rise in urea 59.9 ± 17.6 mg/dL, serum creatinine 1.56 ± 0.97 mg/dL and Cystatin C 199 ± 113 ng/ml as compared to control have serum NGAL 76.31 ± 26.34 ng/ml, urea 22.3 ± 5.7 mg/dL, serum creatinine 0.75 ± 0.14 mg/dL and Cystatin C 76 ± 17 ng/ml (P value <0.05). Conclusion: Serum NGAL closely correlates with serum Cystatin C, creatinine, and eGFR, and serve as a potential early and sensitive marker of impaired kidney function/ kidney injury.


Circulation ◽  
2021 ◽  
Vol 143 (Suppl_1) ◽  
Author(s):  
Changwei Li ◽  
Michael Francis ◽  
Adrianna Westbrook ◽  
Ruiyuan Zhang ◽  
Ye Shen ◽  
...  

Introduction: Most genetic variants for chronic kidney disease (CKD) have been identified in non-coding regions, with functional roles that are difficult to interpret. Hypothesis: A whole exome sequencing study focusing on coding variants will reveal novel mechanisms of kidney function and CKD. Methods: We performed whole exome sequencing analyses of cystatin C among 29,789 UK Biobank (UKB) participants with further confirmation among 4,297 white and 607 African American participants of the Health and Retirement Study (HRS). Conditional analyses for loci achieving exome-wide significance ( P <3.5х10 -7 ) were conducted in UKB using both the exome (n=29,789) and imputed GWAS data (n=295,122). Genomic findings were tested for relevance to baseline estimated glomerular filtration rate (eGFR) and stringently adjudicated CKD progression events among participants of the Chronic Renal Insufficiency Cohort (CRIC) by race and smoking status, using a base model and a full model ( Table ). Results: We identified a common missense variant, CST9 rs2983640, in a previously reported locus ( CST3 intron rs13038305), of which the minor G allele was associated with lower serum cystatin C level (UKB: beta=-0.03 mg/L, P =7.64х10 -92 ; HRS whites: beta=-0.05 mg/L, P =4.71х10 -6 ; HRS African Americans: beta=-0.03 mg/L, P =0.64; and multi-ethnic meta-analysis beta=-0.03 mg/L, P =2.46х10 -91 ). After controlling for the CST3 variant, the G allele was associated with higher cystatin C level (UKB exome: beta=0.003 mg/L, P =0.04; UKB GWAS: beta=0.003 mg/L, P =1.47х10 -10 ). Similar associations were identified in white CRIC participants (direct effect: beta=-0.05 mg/L, P =0.005; conditional effect: beta=0.004 mg/L, P =0.86). The CST9 rs2983640 G allele was associated with lower baseline eGFR (base model beta=-0.33 ml/min/1.73 m 2 , P =1.98х10 -6 ) and higher hazard of developing CKD progression independent of the reported CST3 variant ( Table ). Conclusions: We identified a novel missense variant influencing cystatin C level and CKD progression.


Circulation ◽  
2014 ◽  
Vol 129 (suppl_1) ◽  
Author(s):  
Casey M Rebholz ◽  
Kunihiro Matsushita ◽  
Elizabeth Selvin ◽  
Morgan E Grams ◽  
Josef Coresh

Introduction: Chronic kidney disease (CKD) progression assessed by estimated GFR from creatinine (eGFR-Cr) is a risk factor for cardiovascular disease and end-stage renal disease (ESRD) and has been proposed as a surrogate endpoint for clinical trials. It is unclear if CKD progression assessed by change in different filtration markers has similar risk associations with ESRD. Hypothesis: We hypothesized that percent change in novel kidney filtration markers (β 2 -microglobulin and cystatin C) over a 6-year period would be independently associated with increased risk of ESRD during 15 years of follow-up, similar to the risk seen with change in eGFR-Cr. Methods: We conducted prospective analyses of the ARIC study (N=9,703). β 2 -microglobulin, cystatin C, and creatinine were measured at study visits 1 (1990-92) and 2 (1996-98). Incident ESRD (kidney dialysis or transplant) was defined as entry into the U.S. Renal Data System registry between study visit 2 and September 30, 2011. Cox proportional hazards regression was used to estimate the association between percent change in filtration marker and incident ESRD, adjusting for demographics, kidney disease risk factors, and 1 st measurement of the filtration marker. Results: During a median follow-up of 13.1 years, there were 142 incident ESRD cases. Median eGFR-Cr was 97.3 mL/min/1.73 m 2 at 1 st measurement and 89.0 mL/min/1.73 m 2 at 2 nd measurement. Percent change in eGFR-Cr was moderately correlated with percent change in the inverse of β 2 -microglobulin (r = 0.34) and the inverse of cystatin C (r = 0.36). Progression of CKD (10-25% and >25% decline in filtration function) was associated with increased ESRD risk, with novel markers (β 2 -microglobulin, cystatin C) showing an association at least as strong as the creatinine and eGFR-Cr estimates (Table). Conclusions: CKD progression assessed using novel filtration markers is independently associated with ESRD risk, suggesting the potential utility of measuring change in β 2 -microglobulin and cystatin C in clinical trials.


Stroke ◽  
2019 ◽  
Vol 50 (Suppl_1) ◽  
Author(s):  
Eung-joon Lee ◽  
Han-yeong Jeong ◽  
Heejung Mo ◽  
Chan-hyuk Lee ◽  
Min Kyoung Kang ◽  
...  

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