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[Study on the application of number fluorescence density in detecting autoantibodies titer]
Xiaodong Peng1, Ruiwei Zhang, Lanlan Wang
1Laboratory of Clinical Immunology, West China Hospital of Sichuan University, Chengdu 610041.
Insights
A new number fluorescence density analysis assay offers an objective, economical, and simple method for detecting antinuclear antibody (ANA) titers. This method shows high consistency for common ANA patterns, making it a practical alternative to traditional serum dilution assays.
Area of Science:
- Immunology
- Medical Diagnostics
- Biotechnology
Background:
- Antinuclear antibody (ANA) testing is crucial for diagnosing autoimmune diseases.
- Traditional serum dilution assays for ANA titer determination can be time-consuming and subjective.
- Developing objective and efficient methods for ANA titer detection is essential for clinical practice.
Purpose of the Study:
- To evaluate a novel number fluorescence density analysis assay for antinuclear antibody (ANA) titer detection.
- To establish the correlation between number fluorescence density and traditional serum diluted titers.
- To compare the consistency rates of the new assay with the conventional method across different ANA patterns.
Main Methods:
- Selected optimal camera exposure time (4 seconds) for number fluorescence density analysis.
- Utilized a Spot 32 camera system and ipwin32 software for quantitative fluorescence density measurement.
- Analyzed 4,140 ANA-positive sera to correlate fluorescence density with serum dilution titers and compared consistency rates with conventional methods.
Main Results:
- Established specific number fluorescence density ranges corresponding to serum dilution titers (e.g., 29-50 density vs. 1:100 titer).
- Achieved a high overall consistency rate of 89.4% between the number fluorescence density assay and serum dilution assay.
- Demonstrated excellent consistency for speckled (98.9%), homogenous (99.5%), and mixed (99.8%) ANA patterns, with lower rates for nucleolar and centromere patterns.
Conclusions:
- The number fluorescence density analysis assay is a viable, objective, and cost-effective alternative for determining titers of common ANA patterns (speckled, homogenous, mixed).
- This novel assay simplifies ANA titer detection compared to traditional serum dilution methods.
- The assay's objectivity and efficiency enhance its practical utility in clinical diagnostics for autoimmune diseases.
Abstract:
This study was firstly conducted to detect antinuclear antibody(ANA) titer by using number influorescence density analysis assay instead of serum diluted assay. The best camera explore time was selected. Then 4,140 ANA positive sera were detected to determine the relationship between number influorescence density (detected by number camera system Spot 32 and computer analysis software ipwin32) and serum diluted titer. The consistent rates in different ANA patterns used by the two methods were compared. 4 seconds was found to be the best explore time and the relationship between number influorescence density and serum diluted titer was 29-50 vs 1:100, 51-85 vs 1:320, 86-175 vs 1:1000, 176-215 vs 1:3200, 216-237 vs 1:10,000. According to this standard we detected 3140 ANA positive sera by use of the two methods and observed a total consistency rate of 89.4%. The consistency rates of three ANA patterns including speckled, homogenous, mixture of speckled and homogenous were as high as 98.9%, 99.5%, 99.8% respectively. The lower consistency rate patterns included nucleolar (5.3%), centromere (1.8%), ribosome(12.6%) and other special patterns(0%). For practical purpose, number influorescence density analysis assay can be used in detecting the three main ANA patterns (speckled, homogeneous, mixture of speckled and homogenous) titer instead of serum diluted assay. The number influorescence density analysis assay is more objective, economical and simple than the serum diluted assay.