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Identification of genotype Trichinella spiralis using polymerase chain reaction
I.M. Odoyevskaya
PhD in biological sciences
I.I. Benediktov
doctor of biological sciences
V.V. Aseev
PhD in biological sciences
N.V. Hilyuta
postgraduate
All-Russian Scientific Research Institute of Helminthology
named after K.I. Skryabin, 117218, Moscow, B. Cheremushkinskaya., 28,
-mail: odoevskayaim@rambler.ru
L.A. Bukina
PhD in biological sciences
Vyatka State Agricultural Academy, 610117, Kirov, Oktyabrsky pr., 133,
e-mail: l.bukina5@gmail.com
(Approved by section «Invasive diseases in animals», Division of Veterinary Medicine of RAAS on 23rd
of September 2011, Protocol No 3)
Methods for interspecific and intraspecific differentiation of helminthes, identification of genotype Trichinella spiralis using polymerase chain reaction are developed. These methods detect DNA loci specific for genome T. spiralis and enables to determine connection between natural and synanthropic trichinellosis in different biocenosis. Methods are based on interaction of synthesized oligonucleotide primers with DNA loci typical for this parasite genotype. The amplified fragment length is increasing. Electrophoresis of amplification products enables to visualize the DNA fragments and find their molecular mass. The list of equipment and reagents is presented, the process of preparation of clinical trial materials, release of genomic DNA, preparation of reaction mixture for polymerase chain reaction, amplification and electrophoresis modes are described.
Keywords: Trichinella spiralis, method of polymerase chain reaction, differentiation, DNA.