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Original Article

First Evaluation of an Outbreak of Bovine Babesiosis and Anaplasmosis in Southern Brazil Using Multiplex PCR

Mariana Feltrin Canever1, Luisa Lemos Vieira1, Carolina Reck1, Luisa Richter2, Luiz Claudio Miletti1
The Korean Journal of Parasitology 2014;52(5):507-511.
Published online: October 22, 2014

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Corresponding author (lcmilett@yahoo.com.br)
• Received: April 24, 2014   • Revised: August 5, 2014   • Accepted: August 18, 2014

© 2014, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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First Evaluation of an Outbreak of Bovine Babesiosis and Anaplasmosis in Southern Brazil Using Multiplex PCR
Korean J Parasitol. 2014;52(5):507-511.   Published online October 22, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.5.507
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First Evaluation of an Outbreak of Bovine Babesiosis and Anaplasmosis in Southern Brazil Using Multiplex PCR
Korean J Parasitol. 2014;52(5):507-511.   Published online October 22, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.5.507
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First Evaluation of an Outbreak of Bovine Babesiosis and Anaplasmosis in Southern Brazil Using Multiplex PCR
Image
Fig. 1 Analysis of mPCR products by agarose gel electrophoresis. M, 100 bp molecular size marker; lanes 1, 8, and 9, positive mPCR controls; lane 6, negative mPCR control (distilled water); lanes 2-3 (B. bigemina/B. bovis), 4 (B. bigemina/A. marginale), and 5 (B. bigemina/A. marginale), template DNA. Arrows indicate 278, 356, and 1,000 base pair amplicons, respectively, for B. bigemina, B. bovis, and A. marginale) generated using primer sets described in materials and methods.

Fig. 1

First Evaluation of an Outbreak of Bovine Babesiosis and Anaplasmosis in Southern Brazil Using Multiplex PCR
Pathogen Primer Sequence (5´-3´) Amplicon size (bp) References Babesia bigemina BiIA CAT CTA ATT TCT CTC CAT ACC CCT CC 278 pb [30] BiIB CCT CGG CTT CAA CTC TGA TGC CAA AG Babesia bovis BoF BoR CAC GAG GAA GGA ACT ACC GAT GTT GA 356 pb [31] CCA AGG AGC TTC AAC GTA CGA GGT CA Anaplasma marginale 1773F TGT GCT TAT GGC AGA CAT TTC C 1,000 pb [32] 2957R AAA CCT TGT AGC CCC AAC TTA TCC No. of PCR positive animals
No. of PCR negative animals Total no. of animals Am Bbi Bbo Am/Bbi Am/Bbo Bbi/Bbo Am/Bbi/Bbo Multiplex PCR 3 5 0 12 2 1 3 7 33 Percentage (%) 24.2 (n = 8) 54.6 (n = 18) 21.2 100 Detection of pathogen DNA
Co-detection of pathogen DNA
Number of animals infected/ number of samples (%)
Number of animals infected/ number of samples (%)
Am Bbi Bbo Am/Bbi Am/Bbo Bbi/Bbo Am/Bbi/Bbo 20/33 (60.6) 21/33 (63.6) 6/33 (18.2) 12/33 (36.4) 2/33 (6.1) 1/33 (3.0) 3/33 (9.1)
Table 1. Oligonucleotide primer sequences used in multiplex PCR
Table 2. Multiplex PCR test on field bovine blood samples

Am, Anaplasma marginale; Bbi, Babesia bigemina; Bbo, Babesia bovis.

Table 3. Blood samples from 33 cattle using specific PCR amplification and number of samples showing simultaneous specific PCR amplification of 2 or 3 of the targeted micro-organisms

Am, Anaplasma marginale; Bbi, Babesia bigemina; Bbo, Babesia bovis.

Table 1.

Table 2.

Table 3.

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