Possibilities of using nested PCR-RFLP for taxonomic identification of L3 larvae of the family Trichostrongylidae, Leiper, 1912
https://doi.org/10.31016/1998-8435-2024-18-3-264-273
Abstract
The purpose of the research is to apply molecular genetic research methods to identify the taxonomic affiliation of gastrointestinal parasitic sheep nematodes of the family Trichostrongylidae using nested PCR followed by the restriction fragment length polymorphism (RFLP) analysis.
Materials and methods. Parasitic nematodes, L3 Strongylata larvae obtained from incubated fecal samples of sheep. The genomic DNA was isolated using a commercial kit for DNA extraction from micro-quantities of tissues (Synthol, Moscow) as per the manufacturer’s guidelines. For DNA amplification, a T-100 Bio-Rad thermal cycler and a commercial Eurogen Master Mix reagent kit were used. The PCR regime was performed according to the WAAVP guidelines, 2006. The restriction endonuclease Rsa I of amplified Trichostrongylidae fragments was performed according to guidelines of the enzyme manufacturer (Sibenzyme, Novosibirsk).
Results and discussion. To determine the taxonomic affiliation of Strongylata larvae isolated after incubation of feces from sheep, molecular genetic studies were performed using nested PCR followed by the restriction fragment length polymorphism (RFLP) analysis. This method makes it possible to identify, with the least effort, the genotypes of three species of Strongylata Haemonchus contortus, Trichostrongylus colubriformis, and Teladorsagia circumcincta at the larval stage.
Keywords
Teladorsagia circumcincta,
Haemonchus contortus,
Trichostrongylus colubriformis,
parasitic nematodes,
Trichostrongylidae,
PCR-RFLP,
restriction enzyme
Supporting agencies: The study was conducted with financial support from the Russian Science Foundation Grant No. 23- 26-00220.
About the Authors
I. A. Pimenov
All-Russian Scientific Research Institute for Fundamental and Applied Parasitology of Animals and Plant – a branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV” (VNIIP – FSC VIEV)
Russian Federation
Russian Federation
Pimenov Ilya A., Postgraduate Student
28, Bolshaya Cheremushkinskaya st., Moscow, 117218
I. M. Odoevskaya
All-Russian Scientific Research Institute for Fundamental and Applied Parasitology of Animals and Plant – a branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV” (VNIIP – FSC VIEV)
Russian Federation
Russian Federation
Odoevskaya Irina M., Candidate of Biological Sciences
28, Bolshaya Cheremushkinskaya st., Moscow, 117218
A. M. Plieva
FSBEI HE Ingush State University
GDR
GDR
Plieva Aishet M., Doctor of Biological Sciences, Corresponding Member of the International Academy of Ecology & Life Protection Sciences
I. B. Zyazikova Ave., Magas, Republic of Ingushetia, 386001
A. I. Varlamova
All-Russian Scientific Research Institute for Fundamental and Applied Parasitology of Animals and Plant – a branch of the Federal State Budget Scientific Institution “Federal Scientific Centre VIEV” (VNIIP – FSC VIEV)
Russian Federation
Russian Federation
Varlamova Anastasiya I., Doctor of Biological Sciences
28, Bolshaya Cheremushkinskaya st., Moscow, 117218
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Review
For citations:
Pimenov I.A., Odoevskaya I.M., Plieva A.M., Varlamova A.I. Possibilities of using nested PCR-RFLP for taxonomic identification of L3 larvae of the family Trichostrongylidae, Leiper, 1912. Russian Journal of Parasitology. 2024;18(3):264-273. (In Russ.) https://doi.org/10.31016/1998-8435-2024-18-3-264-273
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