Multi-center evaluation of a user-friendly lateral flow assay to determine IP-10 and CCL4 levels in blood of TB and non-TB cases in Africa.
OBJECTIVE
Multi-center evaluation of a user-friendly lateral flow test for detection of IP-10 and CCL4 levels in Mycobacterium tuberculosis (Mtb) antigen-stimulated whole blood samples from tuberculosis (TB) suspects.


METHODS
A quantitative lateral flow (LF)-based assay platform was applied to detect chemokines IP-10 and CCL4. Chemokine quantitation was achieved using interference-free, fluorescent up-converting phosphor (UCP) labels. The new assays allowed worldwide shipping and storage without requiring a cold chain and were tested at seven institutes (including Ethiopia, Malawi, The Gambia, South Africa, Uganda and Namibia) employing portable lightweight readers for detection of the UCP label. At each site, clinical samples, confirmed TB and non-TB (i.e. other respiratory diseases (ORD)) cases, were collected and analyzed simultaneously with quality control (QC) human IP-10 or CCL4 standards.


RESULTS
Performance of the UCP-LF assay in Africa using QC standards indicated high robustness allowing quantitative detection between 100 and 100,000pg/mL. The optimized assays allowed successful determination of chemokine levels using 1μL whole blood sample from the locally recruited subjects with TB or ORD.


CONCLUSIONS
This African multi-center trial further demonstrated the applicability of the low-tech and robust UCP-LF platform as a convenient quantitative assay for chemokine detection in whole blood. Ambient shipping and storage of all assay reagents and the availability of lightweight standalone readers were acknowledged as essential requirement for test implementation in particular in remote and resource-limited settings.
Department of Molecular Cell Biology, Leiden University Medical Center, The Netherlands. Electronic address: P.Corstjens@LUMC.NL.
Paul L A M
PL
Corstjens
Department of Molecular Cell Biology, Leiden University Medical Center, The Netherlands.
Elisa M
EM
Tjon Kon Fat
Department of Molecular Cell Biology, Leiden University Medical Center, The Netherlands.
Claudia J
CJ
de Dood
Department of Infectious Diseases, Leiden University Medical Center, The Netherlands.
Jolien J
JJ
van der Ploeg-van Schip
Department of Infectious Diseases, Leiden University Medical Center, The Netherlands.
Kees L M C
KL
Franken
DST/NRF Centre of Excellence for Biomedical Tuberculosis Research and SAMRC Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Stellenbosch University, Cape Town, South Africa.
Novel N
NN
Chegou
Vaccines & Immunity, Medical Research Council Unit, Gambia.
Jayne S
JS
Sutherland
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Rawleigh
R
Howe
Armauer Hansen Research Institute, Addis Ababa, Ethiopia.
Adane
A
Mihret
Ethiopian Health and Nutrition Research Institute, Addis Ababa, Ethiopia.
Desta
D
Kassa
University of Namibia, Namibia.
Marieta
M
van der Vyver
University of Namibia, Namibia.
Jacob
J
Sheehama
Karonga Prevention Study, Malawi.
Felanji
F
Simukonda
Department of Medicine, Makerere University CWRU, Kampala, Uganda.
Harriet
H
Mayanja-Kizza
Department of Infectious Diseases, Leiden University Medical Center, The Netherlands.
Tom H M
TH
Ottenhoff
DST/NRF Centre of Excellence for Biomedical Tuberculosis Research and SAMRC Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Stellenbosch University, Cape Town, South Africa.
Gerhard
G
Walzl
Department of Infectious Diseases, Leiden University Medical Center, The Netherlands.
Annemieke
A
Geluk
D016022:Case-Control Studies; D054407:Chemokine CCL4 / Q000097:blood*; D054357:Chemokine CXCL10 / Q000097:blood*; D006801:Humans; D014376:Tuberculosis / Q000097:blood*
C066507:CCL4 protein, human; C114753:CXCL10 protein, human; D054407:Chemokine CCL4; D054357:Chemokine CXCL10
CCL4; Chemokine; Diagnosis, field-friendly, lateral flow; IP-10; Tuberculosis; Up-converting phosphor, UCP