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A Simple Method to Quantitate IP-10 in Dried Blood and Plasma Spots
- Source :
- PLoS ONE, Vol 7, Iss 6, p e39228 (2012), PLoS ONE
- Publication Year :
- 2012
-
Abstract
- Background Antigen specific release of IP-10 is an established marker for infection with M.tuberculosis. Compared to IFN-γ, IP-10 is released in 100-fold higher concentrations enabling the development of novel assays for detection. Dried blood spots are a convenient sample for high throughput newborn screening. Aim To develop a robust and sensitive ELISA-based assay for IP-10 detection in plasma, dried blood spots (DBS) and dried plasma spots (DPS); to validate the ELISA in clinically relevant samples; and to assess the performance of the assay for detection of Cytomegalovirus (CMV) and M.tuberculosis specific immune responses. Method We raised mice and rat monoclonal antibodies against human IP-10 and developed an ELISA. The assay was validated and applied to the detection of CMV and M.tuberculosis specific responses in 18 patients with immune reactivity towards M.tuberculosis and 32 healthy controls of which 22 had immune reactivity towards CMV and none towards M.tuberculosis. We compared the performance of this new assay to IFN-γ. Results The ELISA was reliable for IP-10 detection in both plasma and filter paper samples. The linear range of the ELISA was 2.5–600 pg/ml. IFN-γ was not readily detectable in DPS samples. IP-10 was stabile in filter paper samples for at least 4 weeks at 37°C. The correlation between IP-10 detected in plasma, DPS and DBS samples was excellent (r2>0.97). Conclusions This newly developed assay is reliable for IP-10 quantification in plasma, DBS and DPS samples from antigen stimulated and non-stimulated whole blood. The filter paper assays enable easy sample acquisition and transport at ambient temperature e.g. via the postal system. The system can potentially simplify diagnostic assays for M.tuberculosis and CMV infection.
- Subjects :
- Bacterial Diseases
CXC CHEMOKINES
Cytomegalovirus
lcsh:Medicine
INFLAMMATORY MARKERS
TUBERCULOSIS INFECTION
Mice
0302 clinical medicine
Blood plasma
Pathology
030212 general & internal medicine
lcsh:Science
Immune Response
Whole blood
0303 health sciences
Multidisciplinary
IFN-GAMMA
biology
Chemistry
Antibodies, Monoclonal
Clinical Laboratory Sciences
3. Good health
Interleukin-10
GAMMA-INDUCIBLE PROTEIN-10
Infectious Diseases
Monoclonal
Cytomegalovirus Infections
Medicine
Cytokines
Antibody
RELEASE ASSAYS
CHRONIC HEPATITIS-C
Research Article
Test Evaluation
medicine.drug_class
education
Enzyme-Linked Immunosorbent Assay
Monoclonal antibody
Specimen Handling
03 medical and health sciences
Antigen
Diagnostic Medicine
medicine
WHOLE-BLOOD
Tuberculosis
Animals
Humans
030304 developmental biology
Chromatography
T-CELL RESPONSES
Filter paper
INTERFERON-GAMMA
lcsh:R
Mycobacterium tuberculosis
Virology
Rats
Linear range
Immune System
Case-Control Studies
biology.protein
Clinical Immunology
lcsh:Q
3111 Biomedicine
Biomarkers
General Pathology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- PLoS ONE, Vol 7, Iss 6, p e39228 (2012), PLoS ONE
- Accession number :
- edsair.doi.dedup.....a266c915e44892f0b233121b5b8a21ff