Development and statistical assessment of a paper-based immunoassay for detection of tumor markers.

Paper-based assays are an attractive low-cost option for clinical chemistry testing, due to characteristics such as short time of analysis, low consumption of samples and reagents, and high portability of assays. However, little attention has been given to the evaluation of the performance of these simple tests, which should include the use of a statistical approach to define the choice of best cut-off value for the test. The choice of the cut-off value impacts on the sensitivity and specificity of the bioassay. Here, we developed a paper-based immunoassay for the detection of the carcinoembryonic antigen (CEA) and performed a statistical assessment to establish the assay's cut-off value using the Youden's J index (68.28 A.U.), what allowed for a gain in sensibility (0.86) and specificity (1.0). We also discuss about the importance of defining a gray zone as a safety margin for test (±12% over the cut-off value), eliminating all false positives and false negatives outcomes and avoiding misleading results. The test accuracy was calculated as the area under the curve (AUC) of the receiver operating characteristic (ROC) curve, presenting a value of 0.97, what classifies this test as highly accurate. We propose here a low-cost method capable of detecting carcinoembryonic antigen (CEA) in human serum samples, highlighting the importance of statistical tools to evaluate a new low-cost diagnostic method.

Evidence of the differentiated structural arrangement of constitutive heterochromatin between two populations of Astyanax scabripinnis (Pisces, Characidae)

The composition of heterochromatin classes along the chromosomes of specimens from two populations of the fish Astyanax scabripinnis was examined using fluorescence banding with GC- and AT-DNA specific fluorochromes and fluorescence in situ hybridization (FISH) with an AT-rich satellite DNA (As51) probe. For the pericentromeric heterochromatin blocks neither GC/AT-DNA specific fluorochromes nor the FISH technique produce any response with chromosomes from either of the populations. On the other hand, the telomeric distal heterochromatin blocks of both populations fluoresced when the FISH technique was applied but showed distinct responses after GC-specific fluorochrome treatments, leading us to propose different structural arrangements of the FISH-positive heterochromatins. Such differences in chromosome banding patterns together with other karyotypic differences suggest differentiation of these populations at taxonomic level.