Development and application of a quantitative real-time PCR for the diagnosis of Surra in water buffaloes

S. Konnai; H. Mekata; C.N. Mingala; N.S. Abes; C.A. Gutierrez; J.R.V. Herrera; A.P. Dargantes; W.H. Witola; L.C. Cruz; N. Inoue

doi:10.1016/j.meegid.2009.01.006

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Development and application of a quantitative real-time PCR for the diagnosis of Surra in water buffaloes

Journal article

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Development and application of a quantitative real-time PCR for the diagnosis of Surra in water buffaloes

S. Konnai, H. Mekata, C.N. Mingala, N.S. Abes, C.A. Gutierrez, J.R.V. Herrera, A.P. Dargantes, W.H. Witola, L.C. Cruz and N. Inoue

Infection, Genetics and Evolution, Vol.9(4), pp.449-452

2009

DOI: https://doi.org/10.1016/j.meegid.2009.01.006

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Abstract

Trypanosoma evansi (T. evansi) causes the disease called Surra in domestic animals, which is of great economic importance in South Asian countries. In order to improve the diagnosis of Surra, we endeavored to develop a real-time PCR assay for the detection and quantification of parasites in water buffaloes using specific primers for the T. evansi Rode Trypanozoon antigen type (RoTat) 1.2 Variable Surface Glycoprotein (VSG) gene, which is a known diverse DNA region in trypanosomes. The quantitative detection limit of the assay was 102 trypanosomes per mL of blood, and the identity of the amplicon was confirmed in all assays by melting curve analysis. To evaluate the clinical applicability of this procedure, detection and estimation of parasitemia in blood samples obtained from water buffaloes and horses were conducted. T. evansi was detected in 17/607 (2.8%) blood samples, with parasitemia levels ranging from >101 to 107 parasites per mL of blood. Interestingly, out of the 17 PCR positive animals, 3 had previously received trypanocidal treatment and 1 had abortion history. These data indicate that real-time PCR for the estimation of putative parasitemia levels is a quantitatively and objectively applicable technique for clinical diagnosis of Surra, and could help to understand disease stage and risk of transmission of T. evansi.

Details

Title: Development and application of a quantitative real-time PCR for the diagnosis of Surra in water buffaloes
Authors/Creators: S. Konnai (Author/Creator) - Hokkaido University
H. Mekata (Author/Creator) - Hokkaido University
C.N. Mingala (Author/Creator) - Hokkaido University
N.S. Abes (Author/Creator) - Central Luzon State University
C.A. Gutierrez (Author/Creator) - Central Luzon State University
J.R.V. Herrera (Author/Creator) - Central Luzon State University
A.P. Dargantes (Author/Creator) - Central Mindanao University
W.H. Witola (Author/Creator) - Hokkaido University
L.C. Cruz (Author/Creator) - Central Luzon State University
N. Inoue (Author/Creator) - Obihiro University of Agriculture and Veterinary Medicine
Publication Details: Infection, Genetics and Evolution, Vol.9(4), pp.449-452
Publisher: Elsevier BV
Identifiers: 991005543313807891
Copyright: 2009 Elsevier B.V.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.261 Parasitology - Trypanosoma & Leishmania; 1.261.596 Trypanosoma Biology
Web Of Science research areas: Infectious Diseases
ESI research areas: Molecular Biology & Genetics