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( c c 1 5 / 3 c ( c 1 . /
Camundongos BALB/c fêmeas, 28-30 dias (nt = 512)
Monoinfecções: 10.000 TS/animal, via IP (n = 16)
Infecções mistas: 5.000 TS de cada clone/animal, via IP (n = 16)
8 animais
Controles não
tratados(INT) Confirmação da infecção: ESF do 4oao
10o d.a.i., se necessário HE no 10od.a.i.
Validação do
critério de cura
Validação do critério de cura
8 animais
Tratadosc/ BZ (IT) 100mg/kg, 20 doses
Mortalidade(n = 6)
1oao 90od.a.i. Mortalidade(n = 6)
1oao 90od.a.i.
Positividade da PCR para o kDNA em tecidos
Extração de DNA (Andrade et al., 1999)
PCR (Gomes et al., 1998 modificada)
Entreo 31oe35od.a.i. – FA (n=2)
Entre120oe125od.a.i. – FC (n=2)
Positividade da PCR para o kDNA em tecidos
Extração de DNA (Andrade et al., 1999)
PCR (Gomes et al., 1998 modificada)
Entreo 31oe35od.a.i. – FA (n=2) Entre120oe125od.a.i. – FC (n=2) Parasitemia (n = 6)
Brener, 1962
A partir do 4od.a.i. Parasitemia (n = 6)
Brener, 1962
A partir do 4od.a.i.
ESF
Do31oao60od.a.i. HE Filardi & Brener, 1987
60od.a.i. (30 dias pós-tratamento)
ELISA Volleret al., 1976
90o-120od.a.i. (2 e 3 meses pós-tratamento)
AATV
Martins-Filhoet al., 1995 modificada
120od.a.i. (3 meses pós-tratamento)
PCR
Gomes et al., 1998 modificada
60od.a.i. (30 dias pós-tratamento)
D e te rm in a ç ã o d o í n d ic e d e c u ra Perfil de
Susceptibilidade ao BZ (n =6)
Perfil de
Susceptibilidade ao BZ (n =6)
Camundongos BALB/c fêmeas, 28-30 dias (nt = 512)
Monoinfecções: 10.000 TS/animal, via IP (n = 16)
Infecções mistas: 5.000 TS de cada clone/animal, via IP (n = 16)
8 animais
Controles não
tratados(INT) Confirmação da infecção: ESF do 4oao
10o d.a.i., se necessário HE no 10od.a.i.
Validação do
critério de cura
Validação do critério de cura
8 animais
Tratadosc/ BZ (IT) 100mg/kg, 20 doses
Mortalidade(n = 6)
1oao 90od.a.i. Mortalidade(n = 6)
1oao 90od.a.i.
Positividade da PCR para o kDNA em tecidos
Extração de DNA (Andrade et al., 1999)
PCR (Gomes et al., 1998 modificada)
Entreo 31oe35od.a.i. – FA (n=2)
Entre120oe125od.a.i. – FC (n=2)
Positividade da PCR para o kDNA em tecidos
Extração de DNA (Andrade et al., 1999)
PCR (Gomes et al., 1998 modificada)
Entreo 31oe35od.a.i. – FA (n=2) Entre120oe125od.a.i. – FC (n=2) Parasitemia (n = 6)
Brener, 1962
A partir do 4od.a.i. Parasitemia (n = 6)
Brener, 1962
A partir do 4od.a.i.
ESF
Do31oao60od.a.i. HE Filardi & Brener, 1987
60od.a.i. (30 dias pós-tratamento)
ELISA Volleret al., 1976
90o-120od.a.i. (2 e 3 meses pós-tratamento)
AATV
Martins-Filhoet al., 1995 modificada
120od.a.i. (3 meses pós-tratamento)
PCR
Gomes et al., 1998 modificada
60od.a.i. (30 dias pós-tratamento)
D e te rm in a ç ã o d o í n d ic e d e c u ra Perfil de
Susceptibilidade ao BZ (n =6)
Perfil de
$&! $ $ # $T $ ! $ # & 9 #$ 6</ " c . ! c c %32%Bc c c c
c c c % . . 7- 9 c c ! 7-& 9 b ' / 3 c
( c 1 . / Análise de Isoenzimas
Ben Abdrrazaket al., 1993
Análise de Isoenzimas
Ben Abdrrazaket al., 1993
Extração Isoenzimática
Kilgour & Goldofrey et al., 2007
Amostras isoladas por HE
60od.a.i. (nt-TNC= 78, nt-INT= 102)
Amostras isoladas por HE
60od.a.i. (nt-TNC= 78, nt-INT= 102)
TNC
Animais Tratados não curados
PCR de Microssatélites
Valadares et al., 2007
PCR de Microssatélites
Valadares et al., 2007
INT
Controles não tratados
Obtenção de massa úmida do T. cruzi
Obtenção de massa úmida do T. cruzi
Extração de DNA
Tibayrenc et al., 1993
Caracterização molecular dos clones do
T. cruzi remanescentes nos animais com
infecções mistas
Caracterização molecular dos clones do
T. cruzi remanescentes nos animais com infecções mistas
Amostras de DNA obtidas do Coração e Músculo Esquelético positivas para o kDNA
~30od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas do Coração e Músculo Esquelético positivas para o kDNA
~30od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas diretamente do sangue positivas para o kDNA
60od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas diretamente do sangue positivas para o kDNA
60od.a.i. (nt-TNC= 105, nt-INT= 139)
Extração Isoenzimática
Kilgour & Goldofrey et al., 2007
Amostras isoladas por HE
60od.a.i. (nt-TNC= 78, nt-INT= 102)
Amostras isoladas por HE
60od.a.i. (nt-TNC= 78, nt-INT= 102)
TNC
Animais Tratados não curados
PCR de Microssatélites
Valadares et al., 2007
PCR de Microssatélites
Valadares et al., 2007
INT
Controles não tratados
Obtenção de massa úmida do T. cruzi
Obtenção de massa úmida do T. cruzi
Extração de DNA
Tibayrenc et al., 1993
Caracterização molecular dos clones do
T. cruzi remanescentes nos animais com
infecções mistas
Caracterização molecular dos clones do
T. cruzi remanescentes nos animais com infecções mistas
Amostras de DNA obtidas do Coração e Músculo Esquelético positivas para o kDNA
~30od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas do Coração e Músculo Esquelético positivas para o kDNA
~30od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas diretamente do sangue positivas para o kDNA
60od.a.i. (nt-TNC= 105, nt-INT= 139)
Amostras de DNA obtidas diretamente do sangue positivas para o kDNA
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