Development of a gold nanoparticle-based PCR assay for detection of pig surfactant protein D

Surfactant, pulmonary-associated protein D, also known as SFTPD or SP-D, is a member of the collectin family of C-type lectins that is synthesized in many tissues including respiratory epithelial cells in the lung, and contains one C-type lectin domain and one collagen-like domain. SP-D participates in the innate response to inhaled microorganisms and organic antigens, and contributes to immune and inflammatory regulation within the lung. SP-D is synthesized and secreted by alveolar and bronchiolar epithelial cells, but is also expressed by epithelial cells lining various exocrine ducts and the mucosa of the gastrointestinal and genitourinary tracts. SP-D specifically interacts with glycoconjugates and other molecules expressed on the surface of macrophages, neutrophils, and lymphocytes. In addition, SP-D binds to specific surfactant-associated lipids and can influence the organization of lipid mixtures containing phosphatidylinositol in vitro. The expression of SP-D is increased in response to many forms of lung injury, and deficient accumulation of appropriately oligomerized SP-D might contribute to the pathogenesis of a variety of lung diseases. To improve the detection efficiency of the of pig SP-D expression, the gold nanoparticle PCR established for detection of SP-D mRNA with the primers designed according to swine SP-D gene sequence. In addition, the CDS region of swine SP-D gene was amplified by PCR and inserted into pMD18-T vector constructed the recombinant plasmid pMD-SP-D. Copyright © 2015 American Scientific Publishers All rights reserved.