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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Development of a multiplex loop-mediated isothermal amplification (mLAMP) method for the simultaneous detection of bovine Babesia parasites
Journal of Microbiological Methods, Volume 71, No. 3, Year 2007
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Description
A loop-mediated isothermal amplification (LAMP) technique has been used as a novel nucleic acid detection method, whereby the target DNA can be amplified with high specificity and sensitivity under an isothermal condition using a set of four specific primers. In this study, we designed two sets of the LAMP primers for rhoptry-associated protein-1 genes of Babesia bovis and B. bigemina, in which a restriction enzyme cleavage site was inserted into two pairs of species-specific primers to construct a multiplex LAMP (mLAMP) method by combining these two sets totaling eight primers. The mLAMP method was distinguishable between B. bovis and B. bigemina, simultaneously, due to the subsequent restriction enzyme analysis. The sensitivities of the mLAMP method were 103 and 105 times higher on the detection limits for B. bovis and B. bigemina, respectively, than those of the classical PCR methods. Of 40 blood samples collected from cattle living in Ghana, 12 and 27% were positively detected by the mLAMP for B. bovis and B. bigemina, respectively. Furthermore, 14 and 23% of 90 blood samples from cattle in Zambia showed mLAMP-positive reactions to B. bovis and B. bigemina, respectively. These findings indicate that this mLAMP method is a new convenient tool for simultaneous detection of the bovine Babesia parasites. © 2007 Elsevier B.V. All rights reserved.
Authors & Co-Authors
Iseki, Hiroshi
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Alhassan, Andy
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Ohta, Naomi
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Thekisoe, Oriel M.M.
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Yokoyama, Naoaki
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Inoue, Noboru I.
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Nambota, Andrew Mubila
Zambia, Lusaka
University of Zambia
Yasuda, Jun
Japan, Morioka
Iwate University
Igarashi, Ikuo
Japan, Obihiro
Obihiro University of Agriculture and Veterinary Medicine
Statistics
Citations: 170
Authors: 9
Affiliations: 3
Identifiers
Doi:
10.1016/j.mimet.2007.09.019
ISSN:
01677012
Research Areas
Genetics And Genomics
Study Locations
Ghana
Zambia