Development of a nanogold slot blot inhibition assay for the detection of antibodies against bovine herpesvirus type 1

Abstract

Bovine herpesvirus type 1 (BoHV-1) is recognized as an important pathogen causing respiratory, reproductive, and neurological disorders in cattle and is associated with economic losses to animal industry. Accurate diagnostic methods are needed for prevention of disease transmission. While the virus neutralization test is considered the gold standard method, it requires maintenance of the virus and cell cultures, which is time consuming and expensive. Serological techniques such as enzyme-linked immunosorbent assay (ELISA) are widely applied, as these are easy to perform and provide quick results. In the present study, a nanogold slot blot inhibition assay was developed for the serological diagnosis of BoHV-1 and compared with standard ELISA and horseradish peroxidase (HRP) slot blot assays. Of 42 serum samples tested by ELISA, 32 (76.2%) were positive and 10 (23.8%), were negative. The sensitivity and specificity of the nanogold slot blot inhibition assay was similar to that observed for ELISA and HRP slot blot assays, and a strong correlation was observed between the tests. Thus, the nanogold slot blot inhibition assay may serve as an efficient and rapid alternative to ELISA in settings, where plate-reading equipment is lacking.

Slot blotting and flow cytometry: two efficient assays for platelet antibody screening among patients with platelet refractoriness

“…Namely, these issues could be the determination of the presence, nature of platelet-reactive antibodies in patients' sera, and also the correlation rate of the first two tests with MAIPA assay. The present study is the first screening comparison of SB and FC for PR detection, but in the previous limited researches, SB was considered a relatively streamlined, sensitive, easily adaptable and applicable assay for detection of plasmodium-infected mosquito specimens, type I Glanzmann thrombasthenia and bovine herpesvirus type 1 [21,22,28].…”

“…However, the analysis conducted using this method is subjective and needs skilled personnel and expensive instrumentation. Another used method, slot blotting (SB), is a molecular technique applied to detect proteins/biomolecules and represents a simplification of the Western blot method with the advantages of rapidity, low cost, simplicity and availability that allows multiple samples to be evaluated simultaneously [21,22]. In this method, the sample is loaded directly on a nitrocellulose membrane, and the blotting procedure is performed.…”