tissue: plasma sample collection time: August 2013 sample collection site: Mae Tan, Tak, Thailand sample source: hospital outpatient (PCD) naturally exposed or unexposed to plasmodium sp. parasites: exposed symptomatic or asymptomatic at sample collection: non-malaria fever age (years): 54 qpcr screening for plasmodium genus: negative qpcr plasmodium species: N/A
Extracted molecule
protein
Extraction protocol
n/a
Label
anti-IgG/Cy3
Label protocol
n/a
Hybridization protocol
Serum samples were diluted 1:200 in Protein Array Blocking buffer (Whatman Inc, Sanford, ME) supplemented with 10% (vol/vol) DH5α E. coli lysate (MCLAB, San Francisco, CA) and incubated on arrays over night at 4°C. Microarray slides were incubated in biotin-conjugated anti-IgG Fc fragment secondary antibody (Jackson ImmunoResearch, West Grove, PA) diluted 1:200 in blocking buffer, and detected by incubation with streptavidin-conjugated SureLight P-3 (Columbia Biosciences, Columbia, MD). The slides were washed and air-dried by brief centrifugation.
Scan protocol
Probed array slides were scanned in a Perkin Elmer ScanArray Express HT at a wavelength of 670 nm, at 95% laser power and 55% PMT. The output RGB TIFF files generated by the scanner were quantitated using ProScanArray Express software (Perkin Elmer, Waltham, MA) with spot-specific background correction.
Description
Sample_375
Data processing
For analysis of antibody binding to Pf or Pv proteins on the microarray, the following steps were taken: i) the median background signal of antibody binding to 24 spots of IVTT reaction without DNA template (no template control, NTC) was calculated for each individual sample; ii) the raw values of antibody binding to Pf and Pv polypeptides were divided by their corresponding median NTC value, generating fold-over-control (FOC) values; iii) FOC values were log2-transformed for data normalization (data presented in Sample data tables).
Common asymptomatic and submicroscopic malaria infections in Western Thailand revealed in longitudinal molecular and serological studies: a challenge to malaria elimination.
