Title: Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) for the identification of <i>Trichinella</i> isolates
Abstract: In the present study, polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis was developed to identify 5 species ( Trichinella spiralis , Trichinella britovi , Trichinella nativa , Trichinella nelsoni and Trichinella pseudospiralis ) and 3 phenotypes of uncertain taxonomic status ( Trichinella T5, T6, and T8). Eleven restriction endonucleases were used to restrict 3 DNA fragments (1) a 2800 bp fragment of the 43 kDa excretory–secretory (E–S) protein gene, (2) a 1250 bp fragment amplified with the primer pair SB147A and (3) a 372 bp fragment amplified with the primer pair SB372A. This RFLP method allows the identification of the 8 Trichinella phenotypes as follows: T. spiralis by the Hinf I or Dde I endonuclease restriction of the 2800 bp fragment; T. nativa by the Rsa I restriction of the 2800 bp fragment, or by the Alu I restriction of the 1250 bp fragment; T. britovi and Trichinella T8 by the Alu I restriction of the 1250 bp fragments, and can be discriminated between them by the Ssp I restriction of the 2800 bp fragment; T. pseudospiralis by the Msp I restriction of the 372 bp fragment; T. nelsoni by the Hha I or Alu I restriction of the 2800 bp fragment; Trichinella T5 by the Hha I restriction of the 2800 bp fragment; Trichinella T6 by the Alu I restriction of the 1250 bp fragment; and Trichinella T8 by the Ssp I or Rsa I restriction of the 2800 bp fragment. This study reveals also an intraspecifies polymorphism in the 2800 bp and 1250 bp fragments for T. britovi , Trichinella T5 and T6.
Publication Year: 1999
Publication Date: 1999-02-01
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
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Cited By Count: 63
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