RESUMO
The purpose of this study was to investigate the genotypes of Giardia lamblia from human and animal feces and their epidemiological and clinical characteristics in Argentina, South America. Seventy isolates, 60 from humans (adults and children), eight from dogs and two from cows were processed by polymerase chain reaction-restriction fragment length polymorphism. Data corresponding to demographic, socio-cultural and environmental variables and presence/absence of signs/symptoms were collected. The triosephosphate isomerase gene was amplified from 43 (71.66%) of the 60 human fecal samples. Among these, 3/43 (6.98%) were genotype AII and 40/43 (93.02%) were genotype B. Assemblage AII was detected in three children who lived together in a shantytown and they were oligosymptomatic and none had diarrhea. This genotype was not found in animals. Genotype B showed a high prevalence in both adults and children. It was also found in polysymptomatic people, many of whom presented diarrhea. It was also found only in one dog. The present study represents the first contribution to the knowledge of G. lamblia genotypes in Argentina.
Assuntos
DNA de Protozoário/genética , Fezes/parasitologia , Giardia lamblia/genética , Giardíase/parasitologia , Triose-Fosfato Isomerase/genética , Adolescente , Adulto , Animais , Argentina , Bovinos , Criança , Pré-Escolar , Cães , Feminino , Genótipo , Giardia lamblia/enzimologia , Giardia lamblia/isolamento & purificação , Giardíase/diagnóstico , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Fatores de RiscoRESUMO
The purpose of this study was to investigate the genotypes of Giardia lamblia from human and animal feces and their epidemiological and clinical characteristics in Argentina, South America. Seventy isolates, 60 from humans (adults and children), eight from dogs and two from cows were processed by polymerase chain reaction-restriction fragment length polymorphism. Data corresponding to demographic, socio-cultural and environmental variables and presence/absence of signs/symptoms were collected. The triosephosphate isomerase gene was amplified from 43 (71.66 percent) of the 60 human fecal samples. Among these, 3/43 (6.98 percent) were genotype AII and 40/43 (93.02 percent) were genotype B. Assemblage AII was detected in three children who lived together in a shantytown and they were oligosymptomatic and none had diarrhea. This genotype was not found in animals. Genotype B showed a high prevalence in both adults and children. It was also found in polysymptomatic people, many of whom presented diarrhea. It was also found only in one dog. The present study represents the first contribution to the knowledge of G. lamblia genotypes in Argentina.
Assuntos
Adolescente , Adulto , Animais , Bovinos , Criança , Pré-Escolar , Cães , Feminino , Humanos , Lactente , Masculino , DNA de Protozoário/genética , Fezes/parasitologia , Giardia lamblia/genética , Giardíase/parasitologia , Triose-Fosfato Isomerase/genética , Argentina , Genótipo , Giardia lamblia/enzimologia , Giardia lamblia/isolamento & purificação , Giardíase/diagnóstico , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Fatores de RiscoRESUMO
Giardia lamblia (syn G. intestinalis, G. duodenalis) is the intestinal protozoan producing non-bacterial diarrhea most prevalent in the world. PCR genotype classification of Giardia in feces depends on the quality and quantity of purified DNA and the removal of a great number of inhibitors. The aim of this study was to adapt a PCR protocol to the amplification of the triosephosphate isomerase (tpi) gene of Giardia lamblia in formalin-fixed feces. The tpi gene of G. lamblia was amplified in 28 of the 34 analyzed samples (82.35%) and the B genotype was obtained in all cases. Two major modifications were implemented to improve the performance of PCR from formolated fecal matter. One of these improvements was the use of polyvinylpyrrolidone (PVP) and the other was the addition of bovine serum albumin (BSA). The PCR protocol used in this study showed an amplification percentage exceeding the values reported by other authors with high sensibility and specificity.
Assuntos
Fezes/parasitologia , Giardia lamblia/genética , Reação em Cadeia da Polimerase , Triose-Fosfato Isomerase/genética , Animais , Humanos , Sensibilidade e EspecificidadeRESUMO
Se evaluó la eficiencia de procedimientos de lisis y tratamientos de extracción de ADN de trofozoítos de Giardia lamblia respecto a la eficiencia de ruptura, cantidad y pureza de ADN, además de los tiempos de procesamiento y costos. Se testearon cinco métodos de lisis (agua destilada y calor; agua destilada, calor y proteinasa K; buffer de lisis D; buffer de lisis E y un kit comercial) y tres métodos de purificación de ADN (fenol:cloroformo: isoamílico; Chelex 100 y un kit comercial). Los datos obtenidos se analizaron estadísticamente. La combinación de buffer de lisis E y Chelex fue un método simple y económico, que produjo alto rendimiento de ADN con baja pureza. Ella técnica comercial fue un método simple, más costoso que produjo bajas cantidades de ADN con un nivel de pureza apropiado para estudios moleculares.
