All TEG specimens were run on Thromboelastograph Analyzer 5000 (Haemoscope Corporation, Braintree, MA, USA).
All specimens were run in batches that included blanks, a standard calibration curve, two spiked specimens, and one duplicate.
The technicians were blinded as to the source of the specimens (cases versus controls) and all specimens were run in a single batch.
Known positive serum specimens obtained from patients with confirmed cholera infections and known negative specimens were run on each test occurrence for each assay.
All specimens were run in batches that included a sample blank, a standard calibration curve, and two spiked specimens each time.
At each step, every well contained 50 μl; specimens were run in duplicate in each experiment, and two or three experiments were performed with each plasma/peptide combination.
Serum specimens were run in batches of 10 plus four quality control samples: reagent blank, matrix blank, matrix blank containing a mixed standard of 15 specific congeners and pesticide components at known values, and a duplicate participant sample.
Seven technical replicates (7% of total specimens) were run on different beadchips and dates and the R correlation coefficient of the replicate pairs ranged from 0.9606 to 0.9907 with a median of 0.9852 (Fig. S1B) indicating little variation.
28 All specimens were run using the Applied Biosystems 7500 Fast Dx Real-Time PCR system (Life Technologies, Carlsbad, CA) utilizing the TaqMan EZ RT-PCR Core Reagent Kit (catalog no. N8080236; Life Technologies) with primers and a FAM-BHQ-labeled probe from Integrated DNA Technologies Coralville, IAA).
Each specimen was run in duplicates in the ALS assay and the mean was used to calculate the results.
