Multiplex Assay Kit for Homocysteine (HCy) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-93 84
EDTA plasma(n=5) 78-103 96
heparin plasma(n=5) 87-94 91
sodium citrate plasma(n=5) 78-93 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Homocysteine (HCy) ,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 Homocysteine (HCy) ,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 Homocysteine (HCy) ,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) 87-94% 99-105% 78-101% 91-99%
EDTA plasma(n=5) 84-98% 82-90% 92-101% 89-98%
heparin plasma(n=5) 85-98% 96-103% 84-97% 85-99%
sodium citrate plasma(n=5) 97-105% 85-101% 99-105% 83-97%

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#:HCy) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Journal of Developmental Origins of Health and Disease Maternal low-quality protein diet exerts sex-specific effects on plasma amino acid profile and alters hepatic expression of methyltransferases in adult rat offspring Pubmed:29582727
The Scientific Journal of Al-Azhar Medical Faculty, Girls Homocysteinemia in relation to anemia in hypothyroid patients Doi: 10.4103/sjamf.sjamf_31_18
Science of The Total Environment Organochlorine pesticides exposure may disturb homocysteine metabolism in pregnant women Pubmed: 31787282
AMERICAN JOURNAL OF TRANSLATIONAL RESEARCH Baicalin rescues hyperglycemia-induced neural tube defects via targeting on retinoic acid signaling Pubmed: 32774702
Journal of Diabetes Research Changes and Risk Factors of Skeletal Muscle Mass and Strength in Patients with Type 2 Diabetes over 60 Years Old: A Cross-Sectional Study from China
Journal of Advances in Medical and Pharmaceutical Sciences Homocysteinaemia in Heart Failure Patients in North Eastern Nigeria
Obstetrik ve Neonatoloji T?p Dergisi KARD?YOVASK¨¹LER S?STEM HASTALIKLARINDA R?SK FAKT?R¨¹ OLAN H?PERHOMOS?STE?NEM?'N?N POL?K?ST?K OVER SENDROMUNDAK? YER?
Frontiers in Nutrition Folate, Vitamin B12, and Homocysteine Levels in Women With Neural Tube Defect-Affected Pregnancy in Addis Ababa, Ethiopia Pubmed:35464038
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