Direct sequencing of the PCR amplified SSU rRNA gene of Entamoeba dispar and the design of primers for rapid differentiation from Entamoeba histolytica.
Parasitology
; 112 ( Pt 4): 363-9, 1996 Apr.
Article
en En
| MEDLINE
| ID: mdl-8935948
ABSTRACT
Since 1993, strains of Entamoeba histolytica sensu lato have been assigned to 2 species on the basis of clinical, biochemical, immunological and genetic evidence the pathogenic strains to E. histolytica sensu stricto, the non-pathogenic strains to Entamoeba dispar. Analysis of the gene encoding for the small subunit ribosomal RNA (SSU rDNA) supports the existence of 2 species. However, while 3 whole SSU rDNA sequences are available in the data bases for E. histolytica, only a partial sequence has been published for E. dispar. Here we report a SSU rDNA sequence for E. dispar. Compared to those of E. histolytica, this sequence shows 1.7% nucleotide substitutions. On the basis of our rDNA data, 2 primers were designed to produce polymerase chain reaction (PCR) amplification from both E. histolytica and E. dispar. Primer specificity for the 2 amoebae was assessed both theoretically against the data bases, and experimentally against a collection of eukaryotic and prokaryotic DNAs. The amplified stretch encompasses a polymorphic Dde I restriction site which allows, after cleavage of the fragment, E. histolytica and E. dispar to be distinguished. The reliability of this method of identification was assessed comparing the results with those based on classic isoenzyme analysis.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Polimorfismo de Longitud del Fragmento de Restricción
/
ADN Ribosómico
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ADN Protozoario
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Entamoeba
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Entamoeba histolytica
Límite:
Animals
Idioma:
En
Revista:
Parasitology
Año:
1996
Tipo del documento:
Article
País de afiliación:
Italia