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GENOTYPE CHARACTERIZATION OF Leishmania (Viannia) braziliensis ISOLATED FROM HUMAN AND CANINE BIOPSIES WITH AMERICAN CUTANEOUS LEISHMANIASIS.

Ferreira LT, Gomes AH, Pereira-Chioccola VL - Rev. Inst. Med. Trop. Sao Paulo (2015 May-Jun)

Bottom Line: The results showed nine distinct patterns.These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo.These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Molecular de Parasitas, Instituto Adolfo Lutz, São Paulo, SP, Brazil.

ABSTRACT

Introduction: American tegumentary leishmaniasis (ATL) can be caused by Leishmania (Viannia) braziliensis complex. The evolution of ATL initially results in lesions and can develop into disseminated or diffuse forms. The genetic diversity of L. (V.) braziliensis in some endemic areas of Brazil has been poorly studied, such as in the state of São Paulo. This study analyzed the genetic diversity of L. (V.) braziliensis isolates collected from patients and dogs with LTA from the state of São Paulo.

Methods: Leishmaniasis diagnosis was determined by PCR. The 132 biopsies were collected in different regions of Sao Paulo State, Brazil (36 municipalities). The genetic characterization of L. (V.) braziliensis isolates was tested by RFLP-PCR using DNA extracted from biopsies. The primer set amplified a specific region of Leishmania internal transcribed spacers of the ribosomal DNA locus.

Results: Of the 132 samples, 52 (40%) were completely genotyped by RFLP-PCR (44 from human patients and eight from dogs). The results showed nine distinct patterns. The majority of the genotyped samples were from Sorocaba (30), and the others were distributed among 14 other municipalities. The first pattern was more frequent (29 samples), followed by pattern 2 (nine samples) and pattern 3 (three samples). Patterns 4, 6, 7, 8 and 9 were composed of two samples each and pattern 5 of one sample.

Conclusion: These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo. These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci.

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Related in: MedlinePlus

Restriction patterns of PCR products digested with HhaI in DNA extracted fromstandard Leishmania strains include the following: L.(V.) guyanensis (MHOM/BR/1975/ M4147) (1), L. (L.)amazonensis (IFLA/BR/1967/PH8) (2), L. (L.)major(MHOM/SU/1973/5- ASKH) (3), L. (L.) infantum(MHOM/BR/1974/PP75) (4), and L. (V.) braziliensis(MHOM/BR/1975/M2903) (5). Digested products were resolved in 2% agarose gelstained with ethidium bromide. MM, 50-bp ladder.
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f01: Restriction patterns of PCR products digested with HhaI in DNA extracted fromstandard Leishmania strains include the following: L.(V.) guyanensis (MHOM/BR/1975/ M4147) (1), L. (L.)amazonensis (IFLA/BR/1967/PH8) (2), L. (L.)major(MHOM/SU/1973/5- ASKH) (3), L. (L.) infantum(MHOM/BR/1974/PP75) (4), and L. (V.) braziliensis(MHOM/BR/1975/M2903) (5). Digested products were resolved in 2% agarose gelstained with ethidium bromide. MM, 50-bp ladder.

Mentions: The first experiments were conducted using the DNA extracted from WHO reference strainsto establish the genotype by RFLP-PCR, using the primer set IR1/IR2 and additionaltreatment with HhaI enzymes.Figure 1 shows therestriction patterns of the six WHO reference strains. L. (V.)guyanensis and L. (V.) braziliensis showed the samerestriction profile. On the other hand, L. (L.) amazonensis, L.(L.) major and L. (L.) infantum had specific restrictionprofiles.


GENOTYPE CHARACTERIZATION OF Leishmania (Viannia) braziliensis ISOLATED FROM HUMAN AND CANINE BIOPSIES WITH AMERICAN CUTANEOUS LEISHMANIASIS.

Ferreira LT, Gomes AH, Pereira-Chioccola VL - Rev. Inst. Med. Trop. Sao Paulo (2015 May-Jun)

Restriction patterns of PCR products digested with HhaI in DNA extracted fromstandard Leishmania strains include the following: L.(V.) guyanensis (MHOM/BR/1975/ M4147) (1), L. (L.)amazonensis (IFLA/BR/1967/PH8) (2), L. (L.)major(MHOM/SU/1973/5- ASKH) (3), L. (L.) infantum(MHOM/BR/1974/PP75) (4), and L. (V.) braziliensis(MHOM/BR/1975/M2903) (5). Digested products were resolved in 2% agarose gelstained with ethidium bromide. MM, 50-bp ladder.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4544252&req=5

f01: Restriction patterns of PCR products digested with HhaI in DNA extracted fromstandard Leishmania strains include the following: L.(V.) guyanensis (MHOM/BR/1975/ M4147) (1), L. (L.)amazonensis (IFLA/BR/1967/PH8) (2), L. (L.)major(MHOM/SU/1973/5- ASKH) (3), L. (L.) infantum(MHOM/BR/1974/PP75) (4), and L. (V.) braziliensis(MHOM/BR/1975/M2903) (5). Digested products were resolved in 2% agarose gelstained with ethidium bromide. MM, 50-bp ladder.
Mentions: The first experiments were conducted using the DNA extracted from WHO reference strainsto establish the genotype by RFLP-PCR, using the primer set IR1/IR2 and additionaltreatment with HhaI enzymes.Figure 1 shows therestriction patterns of the six WHO reference strains. L. (V.)guyanensis and L. (V.) braziliensis showed the samerestriction profile. On the other hand, L. (L.) amazonensis, L.(L.) major and L. (L.) infantum had specific restrictionprofiles.

Bottom Line: The results showed nine distinct patterns.These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo.These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Biologia Molecular de Parasitas, Instituto Adolfo Lutz, São Paulo, SP, Brazil.

ABSTRACT

Introduction: American tegumentary leishmaniasis (ATL) can be caused by Leishmania (Viannia) braziliensis complex. The evolution of ATL initially results in lesions and can develop into disseminated or diffuse forms. The genetic diversity of L. (V.) braziliensis in some endemic areas of Brazil has been poorly studied, such as in the state of São Paulo. This study analyzed the genetic diversity of L. (V.) braziliensis isolates collected from patients and dogs with LTA from the state of São Paulo.

Methods: Leishmaniasis diagnosis was determined by PCR. The 132 biopsies were collected in different regions of Sao Paulo State, Brazil (36 municipalities). The genetic characterization of L. (V.) braziliensis isolates was tested by RFLP-PCR using DNA extracted from biopsies. The primer set amplified a specific region of Leishmania internal transcribed spacers of the ribosomal DNA locus.

Results: Of the 132 samples, 52 (40%) were completely genotyped by RFLP-PCR (44 from human patients and eight from dogs). The results showed nine distinct patterns. The majority of the genotyped samples were from Sorocaba (30), and the others were distributed among 14 other municipalities. The first pattern was more frequent (29 samples), followed by pattern 2 (nine samples) and pattern 3 (three samples). Patterns 4, 6, 7, 8 and 9 were composed of two samples each and pattern 5 of one sample.

Conclusion: These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo. These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci.

Show MeSH
Related in: MedlinePlus