ABSTRACT
Aim: This study aimed to evaluate the mean post-test probability (PTP) of the Maturity-onset diabetes of the young (MODY) calculator in a multiethnic cohort of patients previously diagnosed with type 1 diabetes (T1DM). Materials and methods: The MODY probability calculator proposed by Shields and colleagues (2012) was applied to 117 patients from a T1DM outpatient clinic at a tertiary hospital in Brazil. Additionally, two exons of the HNF1A gene were sequenced in eight patients who hadn't received insulin treatment within six months after the diagnosis. Results: 17.1 % of patients achieved PTP >10 %; 11.1 % achieved PTP >25 % (and all patients >30 %), and 7.7 % achieved PTP >40 %. Among the patients who were selected for genetic sequencing, 100 % presented PTP >30 %, with 66.6 % achieving PTP >40 % and 41.6 % achieving PTP >75 %. These cutoffs are as suggested for the Brazilian population, according to previous investigations. No mutation was observed in the sequenced exons. Conclusion: Considering that only around 10 % of the evaluated cases achieved PTP >30 %, it is highly probable that the most suitable cutoff to select patients for genetic sequencing in a Brazilian cohort of T1DM is higher than the cutoff used in Caucasian populations.
ABSTRACT
The objective of this work was to develop a polymeric structure for a biofiltration unit of domestic effluents through microbiological immobilization, capable of promoting the efficient removal of pollutants, meeting local/national Brazilian standards and/or legislation while providing low environmental impact on their production. Four different structures were tested, namely, polypropylene casings without filling material (TF1); polypropylene casings filled with expanded polystyrene grains (TF2); polypropylene casings, filled with polyurethane foam (TF3); and polypropylene casings, filled with polyvinyl chloride pellets (TF4). A flow of 0.216 m3 d-1 was applied to the system, and the biofilters operated in sequential batches with a hydraulic retention time of 6 h. The efficiency potential of the four immobilization structures was verified regarding biochemical and chemical oxygen demand, total ammoniacal nitrogen and total phosphorus. Microbiological analysis of the formed biofilm, performed with the 16S library sequencing method, with amplification of the 16S rRNA V3 and V3-V4 genomic regions, showed a high diversity of microbiological colonization in the four immobilization structures, with better results and consequently greater community stability in TF2. It is recommended using the filter bed made up of unfilled casings, followed by the one filled with expanded polystyrene grains.
Subject(s)
Sewage , Waste Disposal, Fluid , Sewage/chemistry , Waste Disposal, Fluid/methods , Brazil , Polystyrenes , RNA, Ribosomal, 16S , Polypropylenes , Bioreactors , Biofilms , Nitrogen/chemistryABSTRACT
Milk is an essential food, widely consumed by the population. Brazil is one of the world's largest producers of milk. Milk quality is influenced by several factors in all its stages of production. The aim of this study was to determine the microbiological profile of refrigerated and processed raw bovine milk from industries in Vale do Taquari, state of Rio Grande do Sul, Brazil, using metagenomic analysis. A total of six samples were collected, one of refrigerated raw milk from the tanker truck, one of pasteurized milk and one of milk sterilized by the ultra-high temperature (UHT) process, in each of the industries. The identification of the milk microbiota was performed by sequencing the 16S rRNA gene. The results show that refrigerated raw milk has a greater number of microorganisms, followed by pasteurized milk and sterilized milk, successively. Processed milk showed the presence of beneficial microorganisms such as Streptococcus thermophilus and Streptococcus macedonicus. Nevertheless, even UHT milk showed the presence of microorganisms considered harmful, such as the Bacillus cereus group, Aeromonas dhakensis, Enterobacter bacterium and Acinetobacter haemolyticus. Metagenomics is a valuable tool for the thorough evaluation of the milk microbiota in order to implement the processing stages in industries.
Subject(s)
Sequence Analysis/methods , Milk/microbiology , Microbiota , Brazil , Cooled Foods , Raw Foods/analysisABSTRACT
PURPOSE: This study aimed to analyze the frequency of piroplasmids in the blood of dogs in Rio de Janeiro, compare the performance of microscopic techniques, assess the risk factors associated with infections and also molecularly and morphologically characterize the piroplasmids identified. METHODS: In all, 407 blood samples were collected from dogs between 2018 and 2019. These were subjected to microscopic parasitological techniques for thin and thick smears, stained with Giemsa and using a rapid staining kit. The slides were read under an optical microscope and the protozoa were characterized morphometrically. In addition, the blood samples were subjected to molecular characterization for diagnosing piroplasmid species using primers that amplified the gene 18S rRNA. RESULTS: Piroplasmids were detected in 38 (9.3%) samples. Of these, 33 samples presented nucleotide sequences compatible with Babesia vogeli. Most of the positive samples were young, male, defined breeds dogs that had been attended in clinics in São Gonçalo city. Thrombocytopenia and leukopenia were the hematological alterations more observed in positive samples, but positive samples without alterations were also detected. The sex was the only variable that showed statistical differences. Males dogs being more often infected than females (p < 0.05). The microscope slides mostly showed piriform and oval merozoites measuring greater than 2.5 µm in length, which were compatible with B. vogeli. However, smaller forms were also identified, thus demonstrating the polymorphic nature of this parasite. CONCLUSION: Babesia vogeli was detected in blood samples from dogs in the metropolitan cities of Rio de Janeiro by molecular techniques in different parasite morphotypes.
Subject(s)
Babesia , Babesiosis , Dog Diseases , Animals , Babesia/genetics , Babesiosis/diagnosis , Babesiosis/epidemiology , Brazil/epidemiology , Dog Diseases/diagnosis , Dog Diseases/epidemiology , Dogs , Female , Male , RNA, Ribosomal, 18S/geneticsABSTRACT
Analysis of the temporal, seasonal and demographic distribution of dengue virus (DENV) infections in Barbados was conducted using national surveillance data from a total of 3994 confirmed dengue cases. Diagnosis was confirmed either by DENV-specific real time reverse transcriptase polymerase chain reaction (rRT-PCR), or non-structural protein 1 (NS1) antigen or enzyme linked immunosorbent assay (ELISA) tests; a case fatality rate of 0.4% (10/3994) was observed. The prevalence rate of dengue fever (DF) varied from 27.5 to 453.9 cases per 100,000 population among febrile patients who sought medical attention annually. DF cases occurred throughout the year with low level of transmission observed during the dry season (December to June), then increased transmission during rainy season (July to November) peaking in October. Three major dengue epidemics occurred in Barbados during 2010, 2013 and possibly 2016 with an emerging three-year interval. DF prevalence rate among febrile patients who sought medical attention overall was highest among the 10-19 years old age group. The highest DF hospitalisation prevalence rate was observed in 2013. Multiple serotypes circulated during the study period and Dengue virus serotype 2 (DENV-2) was the most prevalent serotype during 2010, whilst DENV-1 was the most prevalent serotype in 2013. Two DENV-1 strains from the 2013 DENV epidemic were genetically more closely related to South East Asian strains, than Caribbean or South American strains, and represent the first ever sequencing of DENV strains in Barbados. However, the small sample size (n = 2) limits any meaningful conclusions. DF prevalence rates were not significantly different between females and males. Public health planning should consider DENV inter-epidemic periodicity, the current COVID-19 pandemic and similar clinical symptomology between DF and COVID-19. The implementation of routine sequencing of DENV strains to obtain critical data can aid in battling DENV epidemics in Barbados.
ABSTRACT
The aim of this study was to investigate the occurrence of Tritrichomonas foetus in cats in the area surrounding the city of Araçatuba municipality, State of São Paulo, Brazil. Fecal samples from 129 cats were collected by rectal flush technique. It was compared two diagnosis methods, direct examination of feces and PCR. The presence of T. foetus DNA was verified using PCR by amplification of 347-bp fragment from the primers TFR3 and TFR4 and amplicons of positive cases were sequenced. Statistical analyses were performed investigating the associations between T. foetus infection with gender, age, breed, presence of diarrhea and/or history of diarrhea, previous treatment, lifestyle, origin, environment, and co-infection. T. foetus was observed in one sample (n=129) by direct microscopic examination of feces while PCR was positive in five samples (3.9%). Giardia species and Cryptosporidium species co-infection was also observed. Statistical analyses showed no significant associations between T. foetus infection and all listed factors, although most positive cats were asymptomatic and lived in multi-cat household. The isolates of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world. So, the occurrence of T. foetus was confirmed in cats in Araçatuba city (Brazil). This parasite must be considered as a differential diagnosis in cats with diarrhea and also in asymptomatic carriers as source of infection in multi-cat environments.(AU)
O objetivo deste estudo foi investigar a ocorrência de Tritrichomonas foetus em gatos na região do município de Araçatuba, SP, Brasil. Foram coletadas amostras fecais de 129 gatos através da técnica de lavado retal. Dois métodos diagnósticos foram comparados, o exame direto das fezes e a PCR. A presença de DNA de T. foetus foi verificada por meio da PCR através da amplificação de 347 pares de bases a partir dos iniciadores específicos TFR3 e TFR4. Posteriormente, os resultados amplificados das amostras positivas foram sequenciadas. Também foi feita análise estatística a fim de investigar a correlação entre infecção por T. foetus e sexo, idade, raça, presença e/ou histórico de diarreia, tratamento prévio, coinfecção, estilo de vida, origem e tipo de ambiente. O protozoário pôde ser observado em uma amostra através do exame direto das fezes e à PCR foram detectadas cinco amostras positivas (3.9%). Foram detectadas coinfecções por Giardia spp. e Cryptosporidium spp. Não foram observadas correlações entre infecção por T. foetus e todos os fatores listados anteriormente, embora a maioria dos felinos positivos fossem assintomáticos e vivessem em ambientes multigatos. O resultado do sequenciamento genético dos isolados das amostras positivas mostrou 100% de similaridade com outros isolados de felinos no mundo. Assim, a ocorrência de T. foetus foi confirmada em gatos em Araçatuba, São Paulo, Brasil. Sendo assim, o parasito deve considerado como diagnóstico diferencial em gatos com diarreia assim como em portadores assintomáticos como fontes de infecção em ambientes multigatos.(AU)
Subject(s)
Animals , Cats , Protozoan Infections/diagnosis , Tritrichomonas foetus/isolation & purification , Parasitic Diseases/diagnosis , Brazil , Polymerase Chain Reaction/veterinary , Diarrhea/etiologyABSTRACT
The aim of this study was to investigate the occurrence of Tritrichomonas foetus in cats in the area surrounding the city of Araçatuba municipality, State of São Paulo, Brazil. Fecal samples from 129 cats were collected by rectal flush technique. It was compared two diagnosis methods, direct examination of feces and PCR. The presence of T. foetus DNA was verified using PCR by amplification of 347-bp fragment from the primers TFR3 and TFR4 and amplicons of positive cases were sequenced. Statistical analyses were performed investigating the associations between T. foetus infection with gender, age, breed, presence of diarrhea and/or history of diarrhea, previous treatment, lifestyle, origin, environment, and co-infection. T. foetus was observed in one sample (n=129) by direct microscopic examination of feces while PCR was positive in five samples (3.9%). Giardia species and Cryptosporidium species co-infection was also observed. Statistical analyses showed no significant associations between T. foetus infection and all listed factors, although most positive cats were asymptomatic and lived in multi-cat household. The isolates of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world. So, the occurrence of T. foetus was confirmed in cats in Araçatuba city (Brazil). This parasite must be considered as a differential diagnosis in cats with diarrhea and also in asymptomatic carriers as source of infection in multi-cat environments.(AU)
O objetivo deste estudo foi investigar a ocorrência de Tritrichomonas foetus em gatos na região do município de Araçatuba, SP, Brasil. Foram coletadas amostras fecais de 129 gatos através da técnica de lavado retal. Dois métodos diagnósticos foram comparados, o exame direto das fezes e a PCR. A presença de DNA de T. foetus foi verificada por meio da PCR através da amplificação de 347 pares de bases a partir dos iniciadores específicos TFR3 e TFR4. Posteriormente, os resultados amplificados das amostras positivas foram sequenciadas. Também foi feita análise estatística a fim de investigar a correlação entre infecção por T. foetus e sexo, idade, raça, presença e/ou histórico de diarreia, tratamento prévio, coinfecção, estilo de vida, origem e tipo de ambiente. O protozoário pôde ser observado em uma amostra através do exame direto das fezes e à PCR foram detectadas cinco amostras positivas (3.9%). Foram detectadas coinfecções por Giardia spp. e Cryptosporidium spp. Não foram observadas correlações entre infecção por T. foetus e todos os fatores listados anteriormente, embora a maioria dos felinos positivos fossem assintomáticos e vivessem em ambientes multigatos. O resultado do sequenciamento genético dos isolados das amostras positivas mostrou 100% de similaridade com outros isolados de felinos no mundo. Assim, a ocorrência de T. foetus foi confirmada em gatos em Araçatuba, São Paulo, Brasil. Sendo assim, o parasito deve considerado como diagnóstico diferencial em gatos com diarreia assim como em portadores assintomáticos como fontes de infecção em ambientes multigatos.(AU)
Subject(s)
Animals , Cats , Protozoan Infections/diagnosis , Tritrichomonas foetus/isolation & purification , Brazil , Diarrhea/etiology , Parasitic Diseases/diagnosis , Polymerase Chain Reaction/veterinaryABSTRACT
Os carrapatos estão envolvidos em processos biológicos de uma grande variedade de organismos patogênicos. O gênero Amblyomma é o de maior importância médica, com a espécie Amblyomma sculptum Berlese, 1888 envolvida no ciclo de transmissão da febre maculosa brasileira (FMB). Neste estudo, objetivou-se a validação molecular para uma diferenciação na característica morfométrica e no tamanho de idiossoma de larvas de duas espécies de carrapatos, Amblyomma dubitatum Neumann, 1899 e A. sculptum. Larvas não alimentadas foram coletadas em duas áreas de transmissão para FMB, por meio da técnica de armadilha atrativa de CO2. Foram identificadas em nível de espécie por morfometria comparativa, análise molecular por PCR e sequenciamento genômico, com validação pela análise de concordância pelo teste Kappa. As larvas de A. dubitatum apresentaram um comprimento significativamente maior que as larvas de A. sculptum. Embora nenhuma outra espécie do gênero Amblyomma tenha sido testada neste estudo, essa técnica poderá ser utilizada nos locais onde levantamentos acarológicos prévios, baseados nos estádios de ninfa e adultos, indicaram a presença de apenas A. sculptum e A. dubitatum, geralmente mantidos por capivaras. Digno de nota, essa condição é muito comum ao longo das áreas endêmicas para FMB na região Sudeste do Brasil.(AU)
Ticks are involved in biological processes of a wide variety of pathogenic organisms. The genus Amblyomma presents the greatest medical importance, with the species Amblyomma sculptum Berlese, 1888 involved in the transmission cycle of Brazilian Spotted Fever (BSF). In this study, we performed a molecular validation of the morphometric differentiation based on the idiosomal length of the larvae of A. dubitatum and A. sculptum. Unfed larvae were collected in two BSF-transmission areas, using the attractive CO2 trap technique. Larvae were identified at the species level by comparative morphometry, molecular analysis by PCR and genomic sequencing, with validation through agreement analysis by the Kappa test. The larvae of A. dubitatum showed a significantly longer idiosomal length than A. sculptum larvae. Although no other species of the genus Amblyomma has been tested in this study, this technique can be applied to places where previous acarological surveillances based on adult and nymphal ticks stages have indicated the presence of only A. sculptum and A. dubitatum, usually sustained by capybaras. Noteworthy, this condition is very common among many BSF-endemic areas in southeastern Brazil.(AU)
Subject(s)
Animals , Ixodidae/classification , Ixodidae/genetics , Rocky Mountain Spotted Fever , Rodentia/parasitologyABSTRACT
ABSTRACT: The aim of this study was to investigate the occurrence of Tritrichomonas foetus in cats in the area surrounding the city of Araçatuba municipality, State of São Paulo, Brazil. Fecal samples from 129 cats were collected by rectal flush technique. It was compared two diagnosis methods, direct examination of feces and PCR. The presence of T. foetus DNA was verified using PCR by amplification of 347-bp fragment from the primers TFR3 and TFR4 and amplicons of positive cases were sequenced. Statistical analyses were performed investigating the associations between T. foetus infection with gender, age, breed, presence of diarrhea and/or history of diarrhea, previous treatment, lifestyle, origin, environment, and co-infection. T. foetus was observed in one sample (n=129) by direct microscopic examination of feces while PCR was positive in five samples (3.9%). Giardia species and Cryptosporidium species co-infection was also observed. Statistical analyses showed no significant associations between T. foetus infection and all listed factors, although most positive cats were asymptomatic and lived in multi-cat household. The isolates of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world. So, the occurrence of T. foetus was confirmed in cats in Araçatuba city (Brazil). This parasite must be considered as a differential diagnosis in cats with diarrhea and also in asymptomatic carriers as source of infection in multi-cat environments.
RESUMO: O objetivo deste estudo foi investigar a ocorrência de Tritrichomonas foetus em gatos na região do município de Araçatuba, SP, Brasil. Foram coletadas amostras fecais de 129 gatos através da técnica de lavado retal. Dois métodos diagnósticos foram comparados, o exame direto das fezes e a PCR. A presença de DNA de T. foetus foi verificada por meio da PCR através da amplificação de 347 pares de bases a partir dos iniciadores específicos TFR3 e TFR4. Posteriormente, os resultados amplificados das amostras positivas foram sequenciadas. Também foi feita análise estatística a fim de investigar a correlação entre infecção por T. foetus e sexo, idade, raça, presença e/ou histórico de diarreia, tratamento prévio, coinfecção, estilo de vida, origem e tipo de ambiente. O protozoário pôde ser observado em uma amostra através do exame direto das fezes e à PCR foram detectadas cinco amostras positivas (3.9%). Foram detectadas coinfecções por Giardia spp. e Cryptosporidium spp. Não foram observadas correlações entre infecção por T. foetus e todos os fatores listados anteriormente, embora a maioria dos felinos positivos fossem assintomáticos e vivessem em ambientes multigatos. O resultado do sequenciamento genético dos isolados das amostras positivas mostrou 100% de similaridade com outros isolados de felinos no mundo. Assim, a ocorrência de T. foetus foi confirmada em gatos em Araçatuba, São Paulo, Brasil. Sendo assim, o parasito deve considerado como diagnóstico diferencial em gatos com diarreia assim como em portadores assintomáticos como fontes de infecção em ambientes multigatos.
ABSTRACT
Os carrapatos estão envolvidos em processos biológicos de uma grande variedade de organismos patogênicos. O gênero Amblyomma é o de maior importância médica, com a espécie Amblyomma sculptum Berlese, 1888 envolvida no ciclo de transmissão da febre maculosa brasileira (FMB). Neste estudo, objetivou-se a validação molecular para uma diferenciação na característica morfométrica e no tamanho de idiossoma de larvas de duas espécies de carrapatos, Amblyomma dubitatum Neumann, 1899 e A. sculptum. Larvas não alimentadas foram coletadas em duas áreas de transmissão para FMB, por meio da técnica de armadilha atrativa de CO2. Foram identificadas em nível de espécie por morfometria comparativa, análise molecular por PCR e sequenciamento genômico, com validação pela análise de concordância pelo teste Kappa. As larvas de A. dubitatum apresentaram um comprimento significativamente maior que as larvas de A. sculptum. Embora nenhuma outra espécie do gênero Amblyomma tenha sido testada neste estudo, essa técnica poderá ser utilizada nos locais onde levantamentos acarológicos prévios, baseados nos estádios de ninfa e adultos, indicaram a presença de apenas A. sculptum e A. dubitatum, geralmente mantidos por capivaras. Digno de nota, essa condição é muito comum ao longo das áreas endêmicas para FMB na região Sudeste do Brasil.(AU)
Ticks are involved in biological processes of a wide variety of pathogenic organisms. The genus Amblyomma presents the greatest medical importance, with the species Amblyomma sculptum Berlese, 1888 involved in the transmission cycle of Brazilian Spotted Fever (BSF). In this study, we performed a molecular validation of the morphometric differentiation based on the idiosomal length of the larvae of A. dubitatum and A. sculptum. Unfed larvae were collected in two BSF-transmission areas, using the attractive CO2 trap technique. Larvae were identified at the species level by comparative morphometry, molecular analysis by PCR and genomic sequencing, with validation through agreement analysis by the Kappa test. The larvae of A. dubitatum showed a significantly longer idiosomal length than A. sculptum larvae. Although no other species of the genus Amblyomma has been tested in this study, this technique can be applied to places where previous acarological surveillances based on adult and nymphal ticks stages have indicated the presence of only A. sculptum and A. dubitatum, usually sustained by capybaras. Noteworthy, this condition is very common among many BSF-endemic areas in southeastern Brazil.(AU)
Subject(s)
Animals , Ixodidae/classification , Ixodidae/genetics , Rocky Mountain Spotted Fever , Rodentia/parasitologyABSTRACT
Os Influenzavirus podem ser classificados de acordo com suas glicoproteínas externas hemaglutinina (HA) e neuraminidase (NA), ambas apresentando alta variabilidade genética e antigênica. No presente estudo foi realizada análise molecular do gene HA, do vírus influenza A (IA) em amostras colhidas de crianças e lactentes com sintomatologia respiratória atendidas no Hospital Universitário da Universidade de São Paulo (USP), durante os anos de 1995 a 2006. Um total de 3.009 amostras foram analisadas por duplex RT-PCR e 4,38% (n=132) foram positivas, sendo 12,1% (n=16) Influenza B e 87,9% (n=116) IA, das quais 9% (n=9) eram H1N1, 91% (n=91) eram H3N2 e 13,8% (n=16) não foram subtipadas. A região HA1 do gene HA de 39 amostras foi sequenciada e as sequências comparadas com as cepas vacinais e circulantes dos respectivos anos. A região de ligação ao receptor foi conservada em todas as amostras e foram verificadas alterações de aminoácidos principalmente nos sítios antigênicos e arredores. No geral, as cepas vacinais foram compatíveis com as circulantes em São Paulo.
The Influenzavirus can be classified according to their external glycoproteins hemagglutinin (HA) and neuraminidase (NA), both showing high genetic and antigenic variability. In the present study was carried out molecular analysis of the HA gene of influenza A (IA) in samples harvested from children and infants, with respiratory symptoms attended at University Hospital, University of Sao Paulo (USP), during the years 1995 to 2006. A total of 3,009 samples were analyzed by duplex RT-PCR and 4.38% (n = 132) were positive, being 12.1% (n = 16) Influenza B and 87.9% (n = 116) IA, where which 9% (n = 9) were H1N1 and 91% (n = 91) were H3N2 and 13.8% (n = 16) did not subtyped. The HA1 region of HA gene of 39 samples were sequenced and the sequences compared with vaccine strains and circulating strains in those years. The receptor-binding region was conserved in all samples and aminoacid changes were observed mainly in the antigenic sites and surroundings. Overall, the vaccine strains were consistent with those circulating in Sao Paulo.
Subject(s)
Humans , Child , Genetics , Alphainfluenzavirus/genetics , Virology , HemagglutininsABSTRACT
Multiple endocrine neoplasia type 2 (MEN2) is an autosomal dominant inherited tumor syndrome caused by RET proto-oncogene germline mutations (RET). Here we tested the Conformation Sensitive Gel Electrophoresis (CSGE) as a screening method for RET hot-spot mutations. Seven MEN2 families were studied by direct sequencing analysis, CSGE and Single Strand Conformational Polymorphism (SSCP). Using CSGE/SSCP, we were able to detect four out of five types of RET mutations verified by sequencing analysis: Cys620Arg, Cys634Arg, Cys634Tyr, and Met918Thr, furthermore a missense substitution at codon 648 (Val648Ile). RET polymorphisms 691 and 769 were also verified. Data obtained using CSGE/SSCP were fully concordant. We conclude that CSGE showed to be a sensitive, fast, low-cost, and simple procedure to detect RET mutations in codons which are reported as the most prevalent RET variants (~ 95 percent) in large MEN2 series. As to the Val804Met mutation, this method still needs to be optimized.
A neoplasia endócrina múltipla tipo 2 (NEM2) é uma síndrome tumoral herdada por mutações germinativas no proto-oncogene RET (RET). Analisamos a aplicação do método Eletroforese em Gel Sensível à Conformação (CSGE) no rastreamento de mutações hot spots do RET. Sete famílias com NEM2 foram rastreadas pelo seqüenciamento gênico, CSGE e análise do Polimorfismo Conformacional de Cadeia Simples (SSCP). Usando ambas as metodologias de rastreamento, identificamos quatro dos cinco tipos de mutações verificadas pelo seqüenciamento: Cys620Arg, Cys634Arg, Cys634Tyr e Met918Thr, além da variação gênica Val648Ile. Das análises englobando mutações hot spots do RET, 90,6 por cento concordaram com o seqüenciamento genético (incluindo a variação gênica Val648Ile). Polimorfismos nos códons 691 e 769 foram documentados. Os dados obtidos por CSGE/SSCP foram totalmente concordantes. Concluímos que o CSGE revelou ser metodologia sensível, rápida, de fácil execução e baixo custo no rastreamento de mutações nos códons associados à grande maioria (~ 95 por cento) dos pacientes com NEM2.