Multiplex Assay Kit for Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay)

TNFAIP5; TSG-14; Pentaxin-Related Gene,Rapidly Induced By IL-1 Beta; Tumor Necrosis Factor Alpha-Induced Protein 5; Tumor necrosis factor-inducible gene 14 protein

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 95-102 99
heparin plasma(n=5) 92-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Pentraxin 3, Long (PTX3) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-95% 93-102% 97-105% 88-98%
EDTA plasma(n=5) 78-102% 94-101% 82-105% 95-102%
heparin plasma(n=5) 80-99% 79-105% 80-91% 97-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:PTX3) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Mediators of Inflammation Biochemical Analysis of Pentraxin 3 and Fibrinogen Levels in Experimental Periodontitis Model PubMed: 22966213
Ibnosina Journal of Medicine and Biomedical Sciences Effect of Vagus Nerve Stimulation on Focal Transient Cerebral Ischemia and Reperfusion in Adult Male White New Zealand Rabbits Sfu: Source
Hemodialysis International Hemodialysis-induced changes in the blood composition affect function of the endothelium Onlinelibrary: hdi.12148
Hemodial Int. Hemodialysis‐induced changes in the blood composition affect function of the endothelium Pubmed:24612443
Journal of Obstetrics and Gynaecology Research The effects of sunitinib on endometriosis Pubmed:25093747
Gynecol Endocrinol. Exenatide improves ovarian and endometrial injury and preserves ovarian reserve in streptozocin induced diabetic rats Pubmed:25366063
Taiwanese Journal of Obstetrics and Gynecology Ovarian failure in diabetic rat model: Nuclear factor-kappaB, oxidative stress, and pentraxin-3 Pubmed:25510691
Journal of Obstetrics and Gynaecology Research Octreotide protects ovary against ischemia–reperfusion injury in rats: Evaluation of histological and biochemical parameters PubMed: 26223286
J Obstet Gynaecol The long pentraxin-3 is a useful marker for diagnosis of ovarian torsion: An experimental rat model PubMed: 26470829
Neurology Research International Accumulation of α-Synuclein in Cerebellar Purkinje Cells of Diabetic Rats and Its Potential Relationship with Inflammation and Oxidative Stress Markers 5952149
Journal of Diabetes Research Study on Association of Pentraxin 3 and Diabetic Nephropathy in a Rat Model Pubmed:29725602
World Journal of Cardiovascular Diseases The Relationship between Serum Pentraxin-3 Levels and Severity of Coronary Heart Disease paperID:86206
Advances in Medical Sciences Serum levels and gene expression of pentraxin 3 are elevated in COPD Pubmed: 30572222
BioMed Research International Pentraxin 3 Detects Clinically Significant Fibrosis in Patients with Chronic Viral Hepatitis C
Evidence-Based Complementary and Alternative Medicine Jie Du Tong Ye San Prevents N-Nitrosomethylbenzylamine-Induced Esophageal Carcinogenesis via Inhibition of Inflammation and Proliferation
Journal of Interferon and Cytokine Research Study on Serum Pentraxin-3 Levels in Vasculitis with Hypertension Pubmed: 31264920
Scientific Reports The role of CRP and Pentraxin 3 in the prediction of systemic inflammatory response syndrome and death in acute pancreatitis Pubmed: 31798002
J Obstet Gynaecol Losartan ameliorates ovarian ischaemia/reperfusion injury in rats: an experimental study Pubmed: 31955629
J Pediatr Adolesc Gynecol Effects of Methylene Blue on Ovarian TorsionLDetorsion Injury in a Rat Model Pubmed: 32593749
Acta Cirurgica Brasileira Therapeutic effects of vitamin D on acetic acid-induced colitis in rats Pubmed: 32555936
Acta Cardiol Sin Pentraxin 3 as a marker for cardiovascular disease risk in overweight and obese children 33716459
International Journal of Endocrinology Effect of Ileal Transposition (IT) on Angiopoietin-Like Protein-8 (ANGPTL8) and Pentraxin (PTX3) Plasma Level in Sprague-Dawley Rats Fed High-Fat Diet?¡­ 34035808
Nutrients High Fat, High Sugar Diet and DJOS Bariatric Surgery Influence Plasma Levels of Fetuin-B, Growth Differentiation Factor-15, and Pentraxin 3 in Diet-Induced Obese … 34684637
J Obstet Gynaecol Res Protective effects of dichloroacetic acid on endometrial injury and ovarian reserve in an experimental rat model of diabetes mellitus 34595798
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