Measurement of salivary cortisol – effects of replacing polyester with cotton and switching antibody

Stable performance between‐runs are essential in longitudinal studies and when different studies are being compared. However, changes in analytical kits and laboratory material occur and have the potential to threaten analytical stability. In the present case, we examined how salivary cortisol measurements in our laboratory were affected by: 1) changes in the tampon material and 2) changes in the antibody of the analytical kit. In study 1, saliva from healthy subjects (n = 19) was split and spiked to Salivette^® polyester and cotton tampons, respectively, and treated as ordinary samples before being analysed for cortisol using a Spectria RIA kit for cortisol. In study 2, 68 anonymous saliva samples were analysed with the Spectria Cortisol RIA kit both before and after the manufacturer changed the antibody. The change from polyester to cotton tampons reduced the measured concentration of salivary cortisol by 62?%. A difference of 12?% between the two runs with different antibodies could not be attributed to differences in storage or in thawing and freezing of samples. To conclude, both a change in the material of the Salivette used for collecting saliva samples as well as a change of antibody in a kit can have a major impact on measurements, as illustrated here for concentrations of cortisol in saliva. It is therefore recommended always to check that the analysis stays in statistical control in one's own laboratory when changes are made, even if the manufacturer reports that the changes should have no effects.     

Impact of scanning density on spectral domain optical coherence tomography assessments in neovascular age‐related macular degeneration

Purpose: To determine the effect of optical coherence tomography (OCT) B‐scan density on the qualitative assessment of neovascular age‐related macular degeneration (AMD). Methods: Data were collected from 59 patients imaged with Topcon 3D OCT‐1000 (128 B‐scans × 512 A‐scans). Custom software was used to generate less dense subsets of scans: 1/16 (eight B‐scans), 1/8 (16 B‐scans), 1/4 (32 B‐scans) and 1/2 (64 B‐scans). At each B‐scan density, scans were assessed for cystoid spaces, subretinal fluid (SRF), subretinal tissue (SRT) and pigment epithelium detachment (PED). For each sampling density, sensitivity, specificity and predictive values were calculated using the full volume scan (128 B‐scans) as the reference standard. Results: For cystoid spaces, the detection sensitivity was 76.3% at 1/16 density; this rose to 89.5% with a 1/4 density. For SRF, the detection sensitivity was 75.0% at a 1/16 density; this increased to 91.1% with 1/4 density. For PED, even at the lowest sampling density (1/16) the detection sensitivity was 86.4%; this rose to 94.9% at 1/4 density. For SRT, detection sensitivity at a 1/16 density was 64.7% and only rose above 90% with the densest sampling subset (1/2). Conclusions: Use of scanning protocols with reduced sampling densities resulted in decreased detection of key features of neovascular AMD; despite this, a sampling density reduced to 1/4 appeared to allow accurate assessment for most features. Current management of neovascular AMD is dependent on qualitative assessment of OCT images; with the recent proliferation of OCT systems, optimization and standardization of scanning protocols may be of value.     

A systematic review classifies sources of bias and variation in diagnostic test accuracy studies

ObjectiveTo classify the sources of bias and variation and to provide an updated summary of the evidence of the effects of each source of bias and variation.Study Design and SettingWe conducted a systematic review of studies of any design with the main objective of addressing bias or variation in the results of diagnostic accuracy studies. We searched MEDLINE, EMBASE, BIOSIS, the Cochrane Methodology Register, and Database of Abstracts of Reviews of Effects (DARE) from 2001 to October 2011. Citation searches based on three key papers were conducted, and studies from our previous review (search to 2001) were eligible. One reviewer extracted data on the study design, objective, sources of bias and/or variation, and results. A second reviewer checked the extraction.ResultsWe summarized the number of studies providing evidence of an effect arising from each source of bias and variation on the estimates of sensitivity, specificity, and overall accuracy.ConclusionsWe found consistent evidence for the effects of case–control design, observer variability, availability of clinical information, reference standard, partial and differential verification bias, demographic features, and disease prevalence and severity. Effects were generally stronger for sensitivity than for specificity. Evidence for other sources of bias and variation was limited.