1921
Volume 94, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract

For differential detection of , , and , loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential identification, one specimen requires three reaction mixtures containing a primer set of each species separately, which is complex and time consuming and increases the risk of cross-contamination. In this study, we developed a simple differential identification of human species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of , , and labeled with fluorescein isothiocyanate (FITC), digoxigenin (DIG), and tetramethylrhodamine (TAMRA), respectively, and biotin-labeled backward inner primers were used in mLAMP. The mLAMP assay succeeded in specific amplification of each respective target gene in a single tube. Furthermore, the mLAMP product from each species was easily distinguished by dot-ELISA with an antibody specific for FITC, DIG, or TAMRA. The mLAMP assay in combination with dot-ELISA will make identification of human species simpler, easier, and more practical.

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  • Received : 16 Nov 2015
  • Accepted : 01 Mar 2016
  • Published online : 01 Jun 2016

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