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Development and validation of direct dry loop mediated isothermal amplification for diagnosis of Trypanosoma evansi

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Title: Development and validation of direct dry loop mediated isothermal amplification for diagnosis of Trypanosoma evansi
Authors: Salim, Bashir Browse this author
Hayashida, Kyoko Browse this author
Mossaad, Ehab Browse this author
Nakao, Ryo Browse this author →KAKEN DB
Yamagishi, Junya Browse this author →KAKEN DB
Sugimoto, Chihiro Browse this author →KAKEN DB
Keywords: Surra
LAMP
Diagnosis
Camel
Trypanosomal
Issue Date: 30-Aug-2018
Publisher: Elsevier
Journal Title: Veterinary Parasitology
Volume: 260
Start Page: 53
End Page: 57
Publisher DOI: 10.1016/j.vetpar.2018.08.009
PMID: 30197015
Abstract: Non-tsetse transmitted Trypanosoma evansi infection (Surra) is one of the most important diseases of camels in north and east Africa and of buffalo and cattle in Asia. Early, accurate and feasible diagnosis is a crucial step towards the control of Surra. Dry format of loop-mediated isothermal amplification (LAMP) diagnostics for the detection of T. evansi was developed, where the detection limit was determined as to equivalent to one parasite per reaction. The assay was validated by testing blood from 48 camels clinically diagnosed to have Surra, which all tested negative microscopically and revealed 43 (89.6%) to be positive for T. evansi when tested by the dry-LAMP. Furthermore, DNA extracted from a randomly selected subset of 20 of these blood samples were then subjected to RoTat1.2-PCR (TaKara Ex Taq), with 14 matching results, with six that were positive by dry-LAMP and negative by PCR. The kappa value of dry-LAMP applied to direct blood was 0.4211, indicating moderate agreement to RoTat 1.2-PCR. In addition, 103 genomic DNA extracted from camels' blood were tested by both dry-LAMP and RoTat1.2-PCR revealed 67 matching results and 31 positive by dry-LAMP and negative by PCR and a further five positives by PCR and negative by dry-LAMP. This novel dry-LAMP method is more sensitive than conventional PCR, direct (without DNA extraction step), is user friendly and does not require cold chain or highly trained personnel.
Rights: © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
http://creativecommons.org/licenses/by-nc-nd/4.0/
Type: article (author version)
URI: http://hdl.handle.net/2115/75288
Appears in Collections:国際連携研究教育局 : GI-CoRE (Global Institution for Collaborative Research and Education : GI-CoRE) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: Bashir Salim

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