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Original Article

Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran

Leila Danehchin1, Gholamreza Razmi1,2,*, Abolghasem Naghibi1
The Korean Journal of Parasitology 2016;54(1):15-20.
Published online: February 26, 2016

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*Corresponding author (razmi@um.ac.ir)
• Received: October 3, 2015   • Revised: November 30, 2015   • Accepted: December 9, 2015

© 2016, 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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Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran
Korean J Parasitol. 2016;54(1):15-20.   Published online February 26, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.1.15
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Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran
Korean J Parasitol. 2016;54(1):15-20.   Published online February 26, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.1.15
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Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran
Image Image Image Image
Fig. 1. PCR amplification products of Toxoplasma gondii B1 in aborted fetuses brain samples Lane M, molecular weight marker (between 1,000 and 50 bp); 1, positive control (Toxoplasma tachyzoites); 2, negative control (H2O instead of DNA); 3-6, positive samples.
Fig. 2. PCR amplification products of Toxoplasma gondii GRA6 in mice brain samples. Lane M, molecular weight marker (between 1,000 and 100 bp); 1-2, positive control (Toxoplasma tachyzoites); 3, negative control (H2O instead of DNA); 4-5, positive samples.
Fig. 3. PCR-RFLP analysis of GRA6 gene coding region with MseI endonuclease. Lanes M1 and M2 are DNA size markers (between 1,000 and 100 bp). Lane 1, undigested PCR products of GRA6 gene; 2-4, digested PCR products of GRA6 gene for Type 1.
Fig. 4. Phylogenetic analysis of representative Iran isolates (Tgshir1,Tgshir2) and reference strains; Strain RH (Type I), GTI (Type I), ME 49 (type II), M7741 (type III), NED (type III), Beverley (type III), RUB (Atypical), C56 (Atypical), and CASTELLE (Atypical). The tree was constructed using the neighbor-joining method after bootstrapping with 1,000 repetitions.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran
Aborted fetuses no. Organ for bioassay No. of seropositive mice/No. of inoculated mice No. of dead mice Day of death (day) Genotyping by GRA6 marker
1 Brain 3/3 3 30 Type I
2 Brain 2/3 3 30 Type I
3 Brain 2/3 0 42 Type I
4 Brain 1/3 0 42 Type I
5 Brain 2/3 0 42
6 Brain 1/3 2 25
7 Brain 1/3 0 42
8 Brain 2/3 0 42
9 Brain 2/3 1 18
10 Brain 1/3 0 42
11 Brain 1/3 0 42
12 Brain 1/3 0 42
13 Brain 1/3 0 42
14 Brain 2/3 0 42
15 Brain 2/3 0 42 Type I
16 Brain 2/3 0 42
17 Brain 1/3 0 42
18 Brain 2/3 0 42
19 Brain 2/3 0 42
20 Brain 2/3 0 42
Strains Positions of variable nucleotides
41 71 106 162 171 195 304 336 561 575 576 614 638 654 677 687 690 692 702
RH C G C G A C A T A G A G G A C C A C T
GT1 * * * * * * * * * * * * * * * * * * *
Tg shir1 * * * * * * * * * * * * * * * * * * *
Tg shir2 * * * * * * * * * * * * - - - - - - -
M 7741 * * * * * * * * * * * C * * * T * * C
RUB * T * * * T T * C C * * * * * * * * *
CASSTTLE * T * A * * T * C * * * A * * * G A *
C56 * T T A * * T * C * * C * * * * * * *
NED * T T A * * T * C * * C * * * * * * *
ME49 T T * * G * T C * * * * * * * * G A A
BEVERLEY T T * * G * T C * * G * - A C * G A A
Table 1. Results of Bioassay and genotyping of Toxoplasma gondii isolated from ovine aborted fetuses which were T. gondii DNA positive cases by nested PCR
Table 2. Comparison of nucleotide sequences of GRA6 gene coding of T. gondii isolates from Mashhad area, Iran with reference strains

Asterisk (*) indicates the same nucleotides of the sequence of RH, and Dashes (-) indicates deletion sites.

Table 1.

Table 2.

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