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Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
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An efficient method for simultaneously screening for HIV, syphilis, and HCV based on one dried blood spot sample
Jieqiong Ma1, Yanan Ren1, Lin He1
1National HIV/HCV Reference Laboratory, National Center for AIDS/STD Control and Prevention, China CDC, Beijing, China.
Antiviral Research
|April 5, 2020
Summary
This study optimized a dried blood spot (DBS) ELISA for simultaneous HIV, Hepatitis C virus (HCV), and Treponema pallidum (TP) antibody screening. The DBS method showed good performance, offering a promising alternative for infectious disease detection.
Area of Science:
- Infectious Diseases
- Immunology
- Biochemistry
Background:
- Dried blood spot (DBS) sampling offers advantages over plasma, including stability, smaller volume, and easier transport, making it suitable for diagnosing various pathogens like HIV, HBV, and HCV.
- While DBS is ideal for resource-limited settings, standardized procedures for simultaneous detection of multiple antibodies (HIV, HCV, TP) are lacking.
Purpose of the Study:
- To optimize parameters for a multiplexed DBS-based ELISA for simultaneous screening of anti-HIV, anti-HCV, and anti-TP antibodies.
- To evaluate the diagnostic performance of the optimized DBS-ELISA assay using paired plasma and DBS samples.
Main Methods:
- Optimized key parameters including blood volume (70-100 μL), DBS size (whole spot), eluent volume (500 μL), eluent composition (PBS with 1‰ Tween20), elution time (4 h), and temperature (room temperature).
- Evaluated assay performance using 429 paired plasma and DBS samples, with plasma results serving as the reference standard.
Main Results:
- The optimized DBS-ELISA demonstrated high clinical sensitivity (HIV: 88.46%, HCV: 98.32%, TP: 92.19%) and 100% specificity for all analytes.
- Positive predictive values were 100% and negative predictive values exceeded 98% for all tested antibodies.
- Only 9 out of 429 samples showed discordant results, primarily with low signal-to-cutoff ratios in plasma.
Conclusions:
- The developed DBS-based ELISA method is effective for simultaneous screening of anti-HIV, anti-HCV, and anti-TP antibodies.
- DBS sampling presents a viable and promising approach for widespread screening of HIV, HCV, and TP infections, particularly in resource-limited settings.
