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PCR identification of Trypanosoma lewisi, a common parasite of laboratory rats.

Desquesnes M, Ravel S, Cuny G - Kinetoplastid Biol Dis (2002)

Bottom Line: T. lewisi has a stringent species specificity and cannot grow in other rodents such as mice.In the course of a research project at CIRDES, a T. lewisi infection was detected in the rat colony.In this study we evaluated PCR primer sets for their ability to diagnose multiple species of trypanosomes with a single amplification.

View Article: PubMed Central - HTML - PubMed

Affiliation: CIRDES, BP454, 01 Bobo-Dioulasso, Burkina Faso. m.desquesnes@fasonet.bf

ABSTRACT
Trypanosoma (Herpetosoma) lewisi is a trypanosome of the sub-genus Herpetosoma (Stercoraria section), parasite of rats (Rattus rattus and Rattus norvegicus) transmitted by fleas. T. lewisi has a stringent species specificity and cannot grow in other rodents such as mice. Rats are infected principally by oral route, through contamination by flea faeces or ingestion of fleas. Trypanosoma lewisi infections in rat colonies can interfere with research protocols and fleas of wild rats are often the source of such infections. Currently, diagnosis of T. lewisi in rats is performed by microscopic observation of stained blood smears. In the course of a research project at CIRDES, a T. lewisi infection was detected in the rat colony. In this study we evaluated PCR primer sets for their ability to diagnose multiple species of trypanosomes with a single amplification. We show that the use of ITS1 sequence of ribosomal DNA provides an efficient and sensitive assay for detection and identification of T. lewisi infection in rats and recommend the use of this assay for monitoring of T. lewisi infections in rat colonies.

No MeSH data available.


Related in: MedlinePlus

Amplification of rDNA ITS with three sets of primers specific for kinetoplastids: A : KIN1 and KIN2 [6] (ITS1), B : IR1 and IR2 [7] (ITS1-2), and C : TRYP1R and TRYP1S (original primers) (ITS1), with DNA of T. lewisi (from 4 different rat buffy-coats 1, 2, 3, 4) T. vivax (5, 6) and T. brucei (7). MM = molecular marker Super ladder 100®, bands are in 100 bp increments, 100 bp band is visible as the topmost band.
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Figure 6: Amplification of rDNA ITS with three sets of primers specific for kinetoplastids: A : KIN1 and KIN2 [6] (ITS1), B : IR1 and IR2 [7] (ITS1-2), and C : TRYP1R and TRYP1S (original primers) (ITS1), with DNA of T. lewisi (from 4 different rat buffy-coats 1, 2, 3, 4) T. vivax (5, 6) and T. brucei (7). MM = molecular marker Super ladder 100®, bands are in 100 bp increments, 100 bp band is visible as the topmost band.

Mentions: With primers TRYP1R and TRYP1S, a 623 bp product was obtained with T. lewisi. The length of this product clearly differentiates T. lewisi from T. brucei, which gave a 520 bp, T. vivax (310 bp) (fig 6) and the T. congolense types (between 680–750 bp; data not shown). TRYP1S and TRYP1R can detect and identify as well the other livestock trypanosomes, with the same level of specificity as that described for KIN primers [5].


PCR identification of Trypanosoma lewisi, a common parasite of laboratory rats.

Desquesnes M, Ravel S, Cuny G - Kinetoplastid Biol Dis (2002)

Amplification of rDNA ITS with three sets of primers specific for kinetoplastids: A : KIN1 and KIN2 [6] (ITS1), B : IR1 and IR2 [7] (ITS1-2), and C : TRYP1R and TRYP1S (original primers) (ITS1), with DNA of T. lewisi (from 4 different rat buffy-coats 1, 2, 3, 4) T. vivax (5, 6) and T. brucei (7). MM = molecular marker Super ladder 100®, bands are in 100 bp increments, 100 bp band is visible as the topmost band.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC119323&req=5

Figure 6: Amplification of rDNA ITS with three sets of primers specific for kinetoplastids: A : KIN1 and KIN2 [6] (ITS1), B : IR1 and IR2 [7] (ITS1-2), and C : TRYP1R and TRYP1S (original primers) (ITS1), with DNA of T. lewisi (from 4 different rat buffy-coats 1, 2, 3, 4) T. vivax (5, 6) and T. brucei (7). MM = molecular marker Super ladder 100®, bands are in 100 bp increments, 100 bp band is visible as the topmost band.
Mentions: With primers TRYP1R and TRYP1S, a 623 bp product was obtained with T. lewisi. The length of this product clearly differentiates T. lewisi from T. brucei, which gave a 520 bp, T. vivax (310 bp) (fig 6) and the T. congolense types (between 680–750 bp; data not shown). TRYP1S and TRYP1R can detect and identify as well the other livestock trypanosomes, with the same level of specificity as that described for KIN primers [5].

Bottom Line: T. lewisi has a stringent species specificity and cannot grow in other rodents such as mice.In the course of a research project at CIRDES, a T. lewisi infection was detected in the rat colony.In this study we evaluated PCR primer sets for their ability to diagnose multiple species of trypanosomes with a single amplification.

View Article: PubMed Central - HTML - PubMed

Affiliation: CIRDES, BP454, 01 Bobo-Dioulasso, Burkina Faso. m.desquesnes@fasonet.bf

ABSTRACT
Trypanosoma (Herpetosoma) lewisi is a trypanosome of the sub-genus Herpetosoma (Stercoraria section), parasite of rats (Rattus rattus and Rattus norvegicus) transmitted by fleas. T. lewisi has a stringent species specificity and cannot grow in other rodents such as mice. Rats are infected principally by oral route, through contamination by flea faeces or ingestion of fleas. Trypanosoma lewisi infections in rat colonies can interfere with research protocols and fleas of wild rats are often the source of such infections. Currently, diagnosis of T. lewisi in rats is performed by microscopic observation of stained blood smears. In the course of a research project at CIRDES, a T. lewisi infection was detected in the rat colony. In this study we evaluated PCR primer sets for their ability to diagnose multiple species of trypanosomes with a single amplification. We show that the use of ITS1 sequence of ribosomal DNA provides an efficient and sensitive assay for detection and identification of T. lewisi infection in rats and recommend the use of this assay for monitoring of T. lewisi infections in rat colonies.

No MeSH data available.


Related in: MedlinePlus