Simple Identification of Human Taenia Species by Multiplex Loop-Mediated Isothermal Amplification in Combination with Dot Enzyme-Linked Immunosorbent Assay

For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c oxidase subunit 1 gene has been recently developed and shown to be sensitive, specific, and effective. However, to achieve differential id...

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Bibliographic Details
Published in:The American journal of tropical medicine and hygiene 2016-06, Vol.94 (6), p.1318-1323
Main Authors: Nkouawa, Agathe, Sako, Yasuhito, Okamoto, Munehiro, Ito, Akira
Format: Article
Language:English
Subjects:
Animals
Base Sequence
DNA - genetics
Electron Transport Complex IV - genetics
Enzyme-Linked Immunosorbent Assay - methods
Humans
Nucleic Acid Amplification Techniques
Sensitivity and Specificity
Taenia
Taenia - classification
Taenia - genetics
Taenia solium
Taeniasis - parasitology
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Summary:For differential detection of Taenia solium, Taenia saginata, and Taenia asiatica, loop-mediated isothermal amplification (LAMP) assay targeting the cytochrome c 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 Taenia 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 Taenia species using multiplex LAMP (mLAMP) in combination with dot enzyme-linked immunosorbent assay (dot-ELISA). Forward inner primers of T. solium, T. saginata, and T. asiatica 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 Taenia species simpler, easier, and more practical.
ISSN:0002-9637
1476-1645
DOI:10.4269/ajtmh.15-0829