Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool
Aim: One of the main diagnostic problems of conventional polymerase chain reaction (PCR) is indiscrimination of low parasitic loads in soil samples. The aim of this study is to determine the genetic diversity and identification of Toxocara spp. from public areas soil inferred by loop-mediated isothe...
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| Format: | Article |
| Language: | English |
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Veterinary World
2016-12-01
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| Series: | Veterinary World |
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| Online Access: | http://www.veterinaryworld.org/Vol.9/December-2016/23.pdf |
| _version_ | 1819141546293854208 |
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| author | Maryam Ozlati Adel Spotin Abbas Shahbazi Mahmoud Mahami-Oskouei Teimour Hazratian Mohammad Adibpor Ehsan Ahmadpour Afsaneh Dolatkhah Paria Khoshakhlagh |
| author_facet | Maryam Ozlati Adel Spotin Abbas Shahbazi Mahmoud Mahami-Oskouei Teimour Hazratian Mohammad Adibpor Ehsan Ahmadpour Afsaneh Dolatkhah Paria Khoshakhlagh |
| author_sort | Maryam Ozlati |
| collection | DOAJ |
| description | Aim: One of the main diagnostic problems of conventional polymerase chain reaction (PCR) is indiscrimination of low parasitic loads in soil samples. The aim of this study is to determine the genetic diversity and identification of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification (LAMP) assay.
Materials and Methods: A total of 180 soil samples were collected from various streets and public parks of northwest Iran. The DNA of recovered Toxocara eggs were extracted and amplified by PCR and LAMP following ZnSO4 flotation technique. The amplicons of internal transcribed spacer-2 gene were sequenced to reveal the heterogeneity traits of Toxocara spp. In addition, Toxocara canis sequences of southwest Iran were directly retrieved to compare gene flow between two distinct populations.
Results: Toxocara spp. eggs were found in 57, 14 and 77 of soil samples using the microscopy, PCR and LAMP (detection limit 1-3 eggs/200 g soil), respectively. 7.7% of isolates were identified as T. canis by PCR method, while LAMP was able to detect 27.2%, 15.5% and 12.2% as Toxocara cati, T. canis and mixed infections, respectively. The kappa coefficient between LAMP and microscopy indicated a strong agreement (0.765) but indicated a faint agreement among LAMP-PCR (0.203) and PCR-microscopy (0.308) methods. A pairwise fixation index (Fst) as a degree of gene flow was generally low (0.02156) among Toxocara populations of northwest and southwest Iran.
Conclusions: The statistically significant Fst value indicates that the T. canis populations are not genetically well differentiated between northwest and southwest Iran. This shows that here is possibly an epidemiological drift due to the transfer of alleles. The LAMP assay because of its shorter reaction time, more sensitivity, and simultaneous detection of environmental contamination to be appears as valuable field diagnosis compared to PCR. Therefore, the detection of low Toxocara spp. loads from public area soils will help to expand epidemiological understanding of toxocariasis and establishing preventive strategies in resource-limited endemic of Iran. |
| first_indexed | 2024-12-22T11:56:10Z |
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| institution | Directory Open Access Journal |
| issn | 0972-8988 2231-0916 |
| language | English |
| last_indexed | 2024-12-22T11:56:10Z |
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| spelling | doaj.art-5dccba588ee24d4781418d07888bc6c22022-12-21T18:26:45ZengVeterinary WorldVeterinary World0972-89882231-09162016-12-019121471147710.14202/vetworld.2016.1471-1477Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular toolMaryam Ozlati0Adel Spotin1Abbas Shahbazi2Mahmoud Mahami-Oskouei3Teimour Hazratian4Mohammad Adibpor5Ehsan Ahmadpour6Afsaneh Dolatkhah7Paria Khoshakhlagh8Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; maryamomedicals@gmail.comInfectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; adelespotin@gmail.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; shahbazy42@yahoo.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; mmahami@gmail.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; hazratian2222@yahoo.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; adibpourm@yahoo.comDrug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; ehsanahmadpour@gmail.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; afsaneh.dolatkhah@gmail.comDepartment of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran; paria.khoshakhlagh@gmail.comAim: One of the main diagnostic problems of conventional polymerase chain reaction (PCR) is indiscrimination of low parasitic loads in soil samples. The aim of this study is to determine the genetic diversity and identification of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification (LAMP) assay. Materials and Methods: A total of 180 soil samples were collected from various streets and public parks of northwest Iran. The DNA of recovered Toxocara eggs were extracted and amplified by PCR and LAMP following ZnSO4 flotation technique. The amplicons of internal transcribed spacer-2 gene were sequenced to reveal the heterogeneity traits of Toxocara spp. In addition, Toxocara canis sequences of southwest Iran were directly retrieved to compare gene flow between two distinct populations. Results: Toxocara spp. eggs were found in 57, 14 and 77 of soil samples using the microscopy, PCR and LAMP (detection limit 1-3 eggs/200 g soil), respectively. 7.7% of isolates were identified as T. canis by PCR method, while LAMP was able to detect 27.2%, 15.5% and 12.2% as Toxocara cati, T. canis and mixed infections, respectively. The kappa coefficient between LAMP and microscopy indicated a strong agreement (0.765) but indicated a faint agreement among LAMP-PCR (0.203) and PCR-microscopy (0.308) methods. A pairwise fixation index (Fst) as a degree of gene flow was generally low (0.02156) among Toxocara populations of northwest and southwest Iran. Conclusions: The statistically significant Fst value indicates that the T. canis populations are not genetically well differentiated between northwest and southwest Iran. This shows that here is possibly an epidemiological drift due to the transfer of alleles. The LAMP assay because of its shorter reaction time, more sensitivity, and simultaneous detection of environmental contamination to be appears as valuable field diagnosis compared to PCR. Therefore, the detection of low Toxocara spp. loads from public area soils will help to expand epidemiological understanding of toxocariasis and establishing preventive strategies in resource-limited endemic of Iran.http://www.veterinaryworld.org/Vol.9/December-2016/23.pdfgene flowIranloop-mediated isothermal amplificationpolymerase chain reactionsoilToxocara spp. |
| spellingShingle | Maryam Ozlati Adel Spotin Abbas Shahbazi Mahmoud Mahami-Oskouei Teimour Hazratian Mohammad Adibpor Ehsan Ahmadpour Afsaneh Dolatkhah Paria Khoshakhlagh Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool Veterinary World gene flow Iran loop-mediated isothermal amplification polymerase chain reaction soil Toxocara spp. |
| title | Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool |
| title_full | Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool |
| title_fullStr | Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool |
| title_full_unstemmed | Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool |
| title_short | Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool |
| title_sort | genetic variability and discrimination of low doses of toxocara spp from public areas soil inferred by loop mediated isothermal amplification assay as a field friendly molecular tool |
| topic | gene flow Iran loop-mediated isothermal amplification polymerase chain reaction soil Toxocara spp. |
| url | http://www.veterinaryworld.org/Vol.9/December-2016/23.pdf |
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