scholarly journals Detection of C-Reactive Protein Using Histag-HRP Functionalized Nanoconjugate with Signal Amplified Immunoassay

Nanomaterials ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1240
Author(s):  
Mohd Farhan Siddiqui ◽  
Zeeshan Ahmad Khan ◽  
Seungkyung Park

Ultrasensitive detection of biomarkers is highly significant for disease prognosis and public health treatment. Despite wide acceptance in routine laboratory tests, the conventional enzyme-linked immunosorbent assay (ELISA) has been of limited use for early biomarker detection due to insufficient sensitivity and multiple long incubation time. Several nanoprobes have been introduced to circumvent the limitation, however, rapid, simple, and chemical-free nanoprobe synthesis and sensitive detection methods, particularly for ELISA, are still lacking. In this study, we have synthesized a gold nanoprobe, conjugated with multiple 6X-histidine (6X-his) peptide and nickel-horseradish peroxidase (Ni2+-HRP), for enhancing the colorimetric signal in ELISA. The developed nanoprobe has been tested for the detection of immunologically significant C-reactive protein (CRP) in ELISA format. The performance of designed probe is validated by testing standard and serum samples, and the detection limit of 32.0 pg/mL with R2 = 0.98 is confirmed. Furthermore, a comparative analysis of the developed nanoprobe was performed with ELISA developed on conventional guidelines, the proposed immunoassay showed an increase of 12-fold sensitivity for detecting CRP due to the high loading of 6Xhis peptide and binding of multiple Ni2+-HRP on a gold nanoparticle. Additionally, the proposed assay provides a simple, fast, and cost-efficient (not requiring multiple antibodies) detection of CRP with easy nanoprobe synthesis. Moreover, the developed Histag-HRP functionalized nanoconjugate immunoassay is flexible and can be applied to other biomarkers efficiently by using disease specific antibody.

2021 ◽  
pp. 1787-1792
Author(s):  
Israa Salim Musa ◽  
Entsar Jabbar Saheb ◽  
Rasha Hussain Kuba

Atherosclerosis is a condition of the hardening of a blood vessel via the development of plaques around the artery wall which causes the artery to narrow, leading to severe complications. Toxoplasmosis is an opportunistic parasitic infection that causes pathological complications in immunocompromised patients, which lead to increase the burden on the immune system in these patients. This study aims to assess the incidence rate of toxoplasmosis in atherosclerosis patients and its potential to change C - reactive protein (C-RP) and vitamin D3 levels. Serum samples (150) were tested for the positivity of anti-Toxoplasma IgG and IgM antibodies by means of Enzyme-linked immunosorbent assay (ELISA). In addition, C-RP was assessed in all serum samples by means of Latex Fixation Test, while VtD3 was estimated by MiniVidas device. The results revealed that the seroprevalence of toxoplasmosis in atherosclerotic patients was comparatively higher as compared to that in the control group, with significant differences in C-RP and VtD3 levels. These results suggest that the decreased levels of VD3 lead to increase the incidence of T. gondii infection in atherosclerosis patients.


2021 ◽  
Vol 3 (1) ◽  
pp. 55-60
Author(s):  
Mohammed Abbas Fadil ◽  
Raya Ezat Maroof ◽  
Moayed Abbas Fadil

Obesity and severe obesity are increasing serious health problems with an epidemic percentage in most countries. In Sleeve gastrectomy, a part of the stomach structure is removed, limiting its capacity by about two to three. A total of thirty blood samples were collected from patients with obesity and the result was compared with healthy person throughout the time from November 2019 to September 2020. Before sleeve gastrectomy and after more than 6 months of sleeve surgery, the sample was collected from the Medical City/Baghdad Teaching Hospital, the withdrawal was again taken at home to have pre and post sleeve gastrectomy, Patient age ranged between [20-46] years for obese patients and healthy control. Then the serum samples were obtained from patients and control group to screen for C-reactive protein by agglutination method. The result of the present study found that the positivity of CRP in pre-operation is higher than that of post-operative with high significance [P<0.005].


Author(s):  
I Mpofana ◽  
M Nyirenda ◽  
N Abbai

Introduction: This study evaluated the performance of the Alere Afinion™ AS100 analyser for the measurement of C-reactive protein (CRP) levels in a population of older adults from South Africa. Methods: This study was a sub-study of the Sexual Health, HIV infection and comorbidity with non-communicable diseases among Older Persons (SHIOP) study. The median age of SHIOP participants was 61 years (interquartile range 12). Serum samples collected through SHIOP were used to measure CRP levels on the Alere Afinion™ AS100 (Point-of-care) and ABX Pentra 400 (reference method), respectively. Bland–Altman analysis and Lin’s concordance correlation coefficients were used to assess the agreement between the two analysers. Results: A total of 183 serum samples were tested in the study. The Alere Afinion™ AS100 median values for CRP were 9.5 mg/L and 11.5 mg/L in women and men respectively (p = 0.275). The ABX Pentra 400 median levels were lower with 5.6 mg/L and 3.6 mg/L for women and men (p = 0.027), respectively. Bland–Altman analysis and linear regression analysis showed an excellent correlation between the Pentra and Afinion analysers, with a Lin’s concordance correlation coefficient of 0.971. The Alere Afinion™ AS100 was able to correctly classify > 90% (165/183) of the CRP results when compared to the ABX Pentra 400. Conclusion: This study showed that the Alere Afinion™ AS100 had an excellent correlation with a standard laboratory method. However, the Afinion™ AS100 did not correlate well at elevated CRP levels. This may not be clinically significant since the cut-points for CVD risk are at much lower levels.


2020 ◽  
Vol 19 (3) ◽  
pp. 2316
Author(s):  
O. V. Khlynova ◽  
E. A. Shishkina ◽  
N. I. Abgaryan

Aim. To study the association of cytokine status with coronary atherosclerosis severity in patients with myocardial infarction (MI).Material and methods. Between 11.2018 and 07.2019, 92 patients hospitalized with MI in Perm Clinical Cardiology Dispensary were included in the study. The control group consisted of 23 patients with stable coronary artery disease. In addition to the standard examination, enzyme-linked immunosorbent assay was used to determine the levels of interleukins (IL)-6, -10, tumor necrosis factor alpha (TNF-α), C-reactive protein.Results. Significant increase in plasma IL-6, TNF-α and C-reactive protein levels in MI patients compared with the control group. The increase in the concentration of IL-6, TNF-α, as well as the IL-6/IL-10 ratio occurs in proportion to coronary atherosclerosis severity. A direct correlation of Gensini score with IL-6, TNF-α, and IL-6/IL-10 ratio was established.Conclusion. Further study of cytokine profile parameters in MI patients will help a clearer understanding pathogenesis of coronary artery atherosclerosis. An increase in concentrations of IL-6, TNF-α, and IL-6/IL-10 ratio is associated with an increase in coronary atherosclerosis severity and can be used in practice for its prediction.


2012 ◽  
Vol 39 (4) ◽  
pp. 728-734 ◽  
Author(s):  
HYOUN-AH KIM ◽  
JA-YOUNG JEON ◽  
JEONG-MI AN ◽  
BO-RAM KOH ◽  
CHANG-HEE SUH

Objective.C-reactive protein (CRP), S100A8/A9, and procalcitonin have been suggested as markers of infection in patients with systemic lupus erythematosus (SLE). We investigated the clinical significance of these factors for indication of infection in SLE.Methods.Blood samples were prospectively collected from 34 patients with SLE who had bacterial infections and 39 patients with SLE who had disease flares and no evidence of infection. A second set of serum samples was collected after the infections or flares were resolved.Results.CRP levels of SLE patients with infections were higher than those with flares [5.9 mg/dl (IQR 2.42, 10.53) vs 0.06 mg/dl (IQR 0.03, 0.15), p < 0.001] and decreased after the infection was resolved. S100A8/A9 and procalcitonin levels of SLE patients with infection were also higher [4.69 μg/ml (IQR 2.25, 12.07) vs 1.07 (IQR 0.49, 3.05) (p < 0.001) and 0 ng/ml (IQR 0–0.38) vs 0 (0–0) (p < 0.001), respectively]; these levels were also reduced once the infection disappeared. In the receiver-operating characteristics analysis of CRP, S100A8/A9, and procalcitonin, the area under the curve was 0.966 (95% CI 0.925–1.007), 0.732 (95% CI 0.61–0.854), and 0.667 (95% CI 0.534–0.799), respectively. CRP indicated the presence of an infection with a sensitivity of 100% and a specificity of 90%, with a cutoff value of 1.35 mg/dl.Conclusion.Our data suggest that CRP is the most sensitive and specific marker for diagnosing bacterial infections in SLE.


2020 ◽  
Vol 18 ◽  
pp. 205873922094234
Author(s):  
Heng Xue ◽  
Hui Liu ◽  
Liangpu Xu ◽  
Qiaoling Liu ◽  
Bimin Zhuo ◽  
...  

The aim of this study was to investigate the predictive value of peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) with C-reactive protein (CRP) and white blood cell (WBC) count for community-acquired pneumonia (CAP) in infants. A total of 84 hospitalized infants with CAP and 69 healthy infants were included in this study. The clinical manifestations and laboratory assay results of infants were recorded. Serum Pin1 level was estimated by enzyme-linked immunosorbent assay. The median serum Pin1 concentration in infants with CAP was significantly higher than that in controls (1.44 vs. 0.21 ng/mL, P < 0.0001). Receiver operating characteristic (ROC) curve analysis showed that the area under the ROC curve (AUC) of the combination Pin1, CRP and WBC (Pin1 + CRP + WBC, 0.943) was higher than Pin1, CRP, WBC alone or the combination of Pin1 and CRP ( P < 0.05). The sensitivity of Pin1 + CRP + WBC (94.0%) was higher than that of Pin1, CRP, WBC alone, or any two combined ( P < 0.05). Pin1 + CRP + WBC also had a high negative predictive value (91.4%). Moreover, serum Pin1 alone had a high specificity (97.0%) and excellent positive predictive value (96.6%) for infants with CAP, which were higher than WBC, Pin1 and WBC in combination, CRP and WBC in combination, and Pin1 + CRP + WBC ( P < 0.05). Therefore, serum Pin 1 was highly expressed in infants with CAP and can singly or in combination with CRP and WBC represent promising novel predictors for infants with CAP.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Leon G. D’Cruz ◽  
Kevin G. McEleney ◽  
Chris Cochrane ◽  
Kyle B. C. Tan ◽  
Priyank Shukla ◽  
...  

AbstractRheumatoid arthritis (RA) is characterised by painful, stiff and swollen joints. RA features sporadic ‘flares’ or inflammatory episodes—mostly occurring outside clinics—where symptoms worsen and plasma C-reactive protein (CRP) becomes elevated. Poor control of inflammation results in higher rates of irreversible joint damage, increased disability, and poorer quality of life. Flares need to be accurately identified and managed. A method comparison study was designed to assess agreement between CRP values obtained by dried blood spot (DBS) versus conventional venepuncture sampling. The ability of a weekly DBS sampling and CRP test regime to detect flare outside the clinic was also assessed. Matched venepuncture and finger lancet DBS samples were collected from n = 100 RA patients with active disease at baseline and 6 weeks (NCT02809547). A subset of n = 30 RA patients submitted weekly DBS samples over the study period. Patient demographics, including self-reported flares were recorded. DBS sample CRP measurements were made by enzyme-linked immunosorbent assay, and venepuncture samples by a reference immunoturbometric assay. Data was compared between sample types by Bland–Altman and weighted Deming regression analyses. Flare detection sensitivity and specificity were compared between ‘minimal’ baseline and 6 week sample CRP data and the ‘continuous’ weekly CRP data. Baseline DBS ELISA assay CRP measures yielded a mean positive bias of 2.693 ± 8.640 (95% limits of agreement − 14.24 to 19.63%), when compared to reference assay data. Deming regression revealed good agreement between the DBS ELISA method and reference assay data, with baseline data slope of 0.978 and intercept -0.153. The specificity of ‘continuous’ area under the curve (AUC) CRP data (72.7%) to identify flares, was greater than ‘minimal’ AUC CRP data (54.5%). This study indicates reasonable agreement between DBS and the reference method, especially at low to mid-range CRP values. Importantly, longitudinal CRP measurement in RA patients helps to clearly identify flare and thus could assist in remote monitoring strategies and may facilitate timely therapeutic intervention.Trial registration: The Remote Arthritis Disease Activity MonitoR (RADAR) study was registered on 22/06/2016 at ClinicalTrials.gov Identifier: NCT02809547. https://clinicaltrials.gov/ct2/show/NCT02809547.


2020 ◽  
Vol 46 (5) ◽  
pp. 1155-1164
Author(s):  
Fu-Chun Zhou ◽  
Joseph W Y Lee ◽  
Qi-Hang Zhang ◽  
Zuo-Li Sun ◽  
Qijing Bo ◽  
...  

Abstract Catatonia is a psychomotor syndrome defined by a constellation of predominantly motor symptoms. The aim of the present study was to determine whether recently admitted psychiatric patients with catatonia exhibited higher serum C-reactive protein (hs-CRP) levels compared to non-catatonic psychiatric patients and healthy controls (HCs). Recently admitted psychiatric patients were screened and evaluated for the catatonia syndrome using the Bush-Francis Catatonia Rating Scale and the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5). The study sample was formed by 150 individuals (39 male and 111 female), including 51 catatonic patients, 55 non-catatonic patients, and 44 HCs. Serum hs-CRP levels were processed with the enzyme-linked immunosorbent assay. Serum levels of creatine kinase (CK), adrenocorticotropic hormone (ACTH), immunoglobulin G (IgG), complement component 3 (C3), and complement component 4 (C4) were also determined. There was a significantly higher percentage of patients with high inflammatory levels (hs-CRP &gt; 3000ng/ml) in the catatonic (43.1%) than in the non-catatonic (14.5%) or HCs group (9.1%) (χ 2 =18.9, P &lt; .001). Logistic regression showed that catatonic patients had significantly higher hs-CRP levels compared to non-catatonic patients even after controlling for other clinical and laboratory variables (OR = 3.52, P = .015, 95% CI 1.28–9.79). Multiple linear regression analysis revealed that log-transformed hs-CRP was independently predicted by body mass index and log-transformed C4, ACTH, and Cortisol in catatonic patients. Findings of the present study suggest that catatonia is specifically linked to a higher level of systemic inflammation, not merely attributable to the overall psychopathology, or alterations in the stress level and complement system.


2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S629-S629
Author(s):  
Supavit Chesdachai ◽  
Nicole Engen ◽  
Joshua Rhein ◽  
Lillian Tugume ◽  
Tadeo kiiza. Kandole ◽  
...  

Abstract Background C-reactive protein (CRP) is an acute-phase protein produced by the liver in response to systemic inflammation. CRP is a helpful surrogate biomarker widely used in various infections, particularly for following the progression and resolution of infection. We aimed to determine the association between baseline CRP level and cryptococcal meningitis outcome. Methods We reviewed 168 prospectively enrolled HIV-infected Ugandans with confirmed first-episode cryptococcal meningitis. Baseline serum samples collected within 5 days from diagnosis had CRP levels measured and categorized into quartiles. We compared baseline serum CRP with 18-week survival using unadjusted time-to-event analysis. Results Of 168 participants, the first quartile of baseline serum CRP was 83.6 mg/L. Baseline CD4 count, HIV viral load, and cerebrospinal fluid results did not differ by quartile. Participants with CRP > 49.5 mg/L more likely presented with Glasgow Coma Scale <15 (P = 0.03). The 18-week mortality rate was 54.8% (46/84) in the highest two quartile CRP groups (49.5 mg/L), 40.5% (17/42) in the mid-range CRP group (29–49.5 mg/L), and 14.3% (6/42) in the low CRP group (<29 mg/L) (P < 0.001) (Figure 1). Conclusion Higher baseline serum CRP is associated with increased mortality in HIV-infected individuals with first-episode cryptococcal meningitis. The serum CRP could be a surrogate marker for undiagnosed co-infections or may reflect immune dysregulation leading to worse outcomes in persons with advanced AIDS and concomitant cryptococcal meningitis. Additional studies investigating more specific inflammatory biomarkers and the longitudinal trend in CRP with effective therapy would be informative. Disclosures All authors: No reported disclosures.


2017 ◽  
Vol 15 (2) ◽  
pp. 120-123
Author(s):  
Takashi Kuribayashi

The half-life ( t1/2) of C-reactive protein (CRP) and its ability to stimulate weak inflammatory responses were investigated in beagle dogs. Four beagle dogs were administered 20 mg/kg indomethacin and blood was collected from the cephalic vein pre-dosing and at 24, 48, 72, 96, 144, 192, 240, 312, and 360 h post-administration. The serum concentrations of CRP were measured by enzyme-linked immunosorbent assay. The serum t1/2 was calculated using the equation 0.693/elimination rate constant. The serum concentration of CRP beyond 192 h post-administration declined to levels in the normal range. The t1/2 was 148.3 h, which is considered to be the essential t1/2 of CRP. The simulation of CRP serum concentrations at arbitrary times using the elimination rate constant obtained in this study became possible.


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