Multiplex Assay Kit for Pepsin (PP) ,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 Pepsin (PP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Pepsin (PP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-105 101
EDTA plasma(n=5) 79-95 91
heparin plasma(n=5) 88-95 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pepsin (PP) ,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 Pepsin (PP) ,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 Pepsin (PP) ,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) 78-103% 97-105% 96-104% 96-103%
EDTA plasma(n=5) 83-91% 93-105% 84-91% 90-101%
heparin plasma(n=5) 95-103% 91-98% 78-94% 78-101%

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#:PP) 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
Radiology and Oncology LEVEL OF PEPSIN AND BILE ACIDS IN THE SALIVA OF PATIENTS WITH GLOTTIS T1 CARCINOMA Szd:Source
Radiology and Oncology Higher levels of total pepsin and bile acids in the saliva as a possible risk factor for early laryngeal cancer Pubmed:Pmc4362607
Clin Otolaryngol Pepsin and bile acids in saliva in patients with laryngopharyngeal reflux – a prospective comparative study PubMed: 25516364
journal of pediatric gastroenterology and nutrition Does Positioning Affect Tracheal Aspiration of Gastric Content in Ventilated Infants? PubMed: 25313850
Journal of Voice Detecting Laryngopharyngeal Reflux by Immunohistochemistry of Pepsin in the Biopsies of Vocal Fold Leukoplakia pubmed:28756936
Sensors (Basel) Label-Free Detection of Salivary Pepsin Using Gold Nanoparticle/Polypyrrole Nanocoral Modified Screen-Printed Electrode Pubmed:29882917
The Tohoku Journal of Experimental Medicine Association between Pepsin in Bronchoalveolar Lavage Fluid and Prognosis of Chronic Fibrosing Interstitial Lung Disease
Respiratory Research Pulmonary aspiration in preschool children with cystic fibrosis Pubmed: 30558606
Tanaffos Association of Bile Acid and Pepsin Micro-aspiration with Chronic Obstructive Pulmonary Disease Exacerbation Pubmed: 31423141
Sensors Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux Pubmed: 31935973
BIOSCIENCE REPORTS Presence of pepsin in laryngeal tissue and saliva in benign and malignant neoplasms Pubmed: 33103719
OTOLARYNGOLOGY-HEAD AND NECK SURGERY Relationship Between Pepsin Expression and Dysplasia Grade in Patients With Vocal Cord Leukoplakia Pubmed: 32692278
American Journal of Otolaryngology Association of pepsin and DNA damage in laryngopharyngeal reflux-related vocal fold polyps Pubmed: 32889371
The relationship between laryngopharyngeal reflux based on pepsin value and clinical characteristics of laryngeal cancer patients
Med Sci Monit The Role of Salivary Pepsin in the Diagnosis of Gastroesophageal Reflux Disease (GERD) Evaluated Using High-Resolution Manometry and 24-Hour?¡­ 33220027
Annals of Esophagus Pepsin properties, structure, and its accurate measurement: a narrative review
Laryngoscope The Impact of Laryngopharyngeal Reflux on Occurrence and Clinical Course of Recurrent Respiratory Papillomatosis 34338331
Pulmonology Bronchoalveolar lavage (BAL) amylase and pepsin levels as potential biomarkers of aspiration pneumonia Pubmed:35715334
Neuromodulation: Technology at the Neural Interface Transcutaneous Electrical Acustimulation Improved the Quality of Life in Patients With Diarrhea-Irritable Bowel Syndrome Pubmed:35088760
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
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