Rodrigo Suzuki | Universidade Federal do ABC (original) (raw)

Papers by Rodrigo Suzuki

Research paper thumbnail of A highly sensitive Leishmania infantum chagasi isolation method from bone marrow and peripheral blood of adults and children

Journal of infection in developing countries, Jan 24, 2016

Research paper thumbnail of Novel Single Hematophagous Insect RNA Detection Method Supports Its Use as Sentinels to Survey Flaviviruses Circulation

Dengue Fever [Working Title], Apr 10, 2020

Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, con... more Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, contribute to emergence and reemergence of arboviruses worldwide, where Flavivirus is the most prevalent, and its continuous monitoring can help in preventive control strategies. Thus, the aim of this study was to detect flavivirus RNA in single hematophagous insects, which are used as sentinels. Total RNA was extracted from six Aedes aegypti stored since 2003 and from 100 Culicidae and collected through CDC trap in a public park of a Brazilian Northwest city of São Paulo State. Flavivirus was detected through RT/PCR targeting 230-250 bp of the RNA polymerase coding sequence (NS5). PCR amplicons were sequenced by Sanger method, used in comparative analysis over Basic Local Alignment Search Tool (BLAST) in GenBank, and subjected to Neighbor-Joining phylogenetic analyses. Efficiency of Flavivirus diagnosis was confirmed by detection of Dengue virus serotype 2 in Ae. aegypti. From the 100 collected insects, 19 were positive for Culex flavivirus (CxFV). NS5 partial sequence phylogenetic analysis clustered all CxFV in one branch separated from vertebrate flaviviruses, being applicable to the identification of Flavivirus species. The dipteran RNA extraction methodology described in this work supports detection of flaviviruses in single insects maintained in 80% ethanol, which can be used to constant arbovirus surveillance.

Research paper thumbnail of A PCR-based strategy for Dombrock screening in Brazilian blood donors reveals a novel allele: the DO*A-WL

Journal of Clinical Laboratory Analysis, 2011

Background: Determination of the molecular basis underlying the antigens in the Dombrock blood gr... more Background: Determination of the molecular basis underlying the antigens in the Dombrock blood group system has shown various rearrangements between the alleles associated with DO Ã A and DO Ã B. Based on this, we employed a PCR-based strategy to screen DO alleles (DO Ã A, DO Ã B, HY Ã 1, HY Ã 2 and JO) in Brazilians. Methods: We tested DNA of 278 Brazilian blood donors by PCR-RFLP on plates of 96 wells to determine the 793A/G (DO Ã A/DO Ã B), 323G/T (HY), 350C/T (JO) and 898C/G (HY Ã 1/HY Ã 2) single nucletide polymorphisms. In order to confirm the results sequence analysis was also performed. Results: When samples of these donors were analyzed, a novel allele combination, the DO Ã A allele (793A and 323G)

Research paper thumbnail of A Highly Sensitive and Specific Conventional Molecular Diagnosis for Leishmania infantum Chagasi Based on a Single Copy Gene

Journal of Molecular Biomarkers & Diagnosis, 2016

Research paper thumbnail of Nucleic Acid-based Diagnosis and Epidemiology of Infectious Diseases

Nucleic Acids - From Basic Aspects to Laboratory Tools, 2016

Research paper thumbnail of HNA-3 gene frequencies in Brazilians and a new polymerase chain reaction-restriction fragment length polymorphism method for HNA-3a/3b genotyping

Transfusion, 2013

BACKGROUND: HNA-3 antigens are the result of a rs2288904 single-nucleotide polymorphism (SNP) in ... more BACKGROUND: HNA-3 antigens are the result of a rs2288904 single-nucleotide polymorphism (SNP) in the CTL2, and the HNA-3a and HNA-3b variants are encoded by a guanine and adenine at Nucleotide Position 461. Anti-HNA-3 are involved in severe transfusionrelated acute lung injury reactions and in neonatal alloimmune neutropenia. Since the distribution of the HNA-3 system was unknown in South Americans, in this study we determined the frequency of the HNA-3 alleles in Brazilians. STUDY DESIGN AND METHODS: DNA of 500 blood donors, 120 Xikrin Amerindians, 74 Japanese individuals, and 124 African Brazilians were genotyped for rs2288904 by a polymerase chain reaction (PCR)restriction fragment length polymorphism assay. The PCR product was digested with enzyme Taq α 1, specific to nucleotide guanine (HNA-3a).

Research paper thumbnail of Dombrock genotyping in Brazilian blood donors reveals different regional frequencies of the HY allele

Revista Brasileira de Hematologia e Hemoterapia, 2013

The frequencies of the DO alleles in Minas Gerais, a southeastern state of Brazil, were determine... more The frequencies of the DO alleles in Minas Gerais, a southeastern state of Brazil, were determined from the genotyping of 270 blood donors. Genotyping involved polymerase chain reaction and restriction fragment length polymorphism analysis to identify the 323G>T, 350C>T, 793A>G, and 898C>G mutations, which are related to the HY, JO, DO*A/DO*B, and DO*A-WL/DO*B-WL alleles, respectively. Moreover, the frequencies of rare HY and JO alleles were statistically compared using the chi-square test with data from another Brazilian region.

Research paper thumbnail of Rural Triatoma rubrovaria from southern Brazil harbors Trypanosoma cruzi of lineage IIc

The American journal of tropical medicine and hygiene, 2008

Triatoma infestans, the main vector of Chagas disease, has nearly been eliminated from Brazil. Ne... more Triatoma infestans, the main vector of Chagas disease, has nearly been eliminated from Brazil. Nevertheless, other triatominae species are involved in the domiciliation process, including Triatoma rubrovaria in Rio Grande do Sul State (RS). Previous studies showed that 1.6% of the T. rubrovaria specimens collected at the rural district of Quaraí, RS, were naturally infected by Trypanosoma cruzi. In this study, five T. cruzi isolates obtained from infected triatomines were characterized molecularly and biologically. Genotyping of the T. cruzi isolates showed that they belong to lineage IIc of T. cruzi (TCIIc). Biological characterization showed miotropism and myositis during acute and chronic phases of infection, respectively. Virulence and mortality rates were variable among isolates. To our knowledge, this study corresponds to the first characterization of T. cruzi isolates from T. rubrovaria and the first description of TCIIc in the sylvatic cycle of T. cruzi from the southern reg...

Research paper thumbnail of Isolation and Characterization of Mayaro Virus from a Human in Acre, Brazil

The American journal of tropical medicine and hygiene, Jan 15, 2014

Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of ... more Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of Mayaro fever, an acute febrile illness often presenting with arthralgic manifestations. The true incidence of MAYV infection is likely grossly underestimated because the symptomatic presentation is very similar to that of dengue fever and other acute febrile tropical diseases. We report the complete genome sequence of a MAYV isolate detected from an Acrelândia patient presenting with fever, chills, and sweating, but with no arthralgia. Results show that this isolate belongs to genotype D and is closely related to Bolivian strains. Our results suggest that the Acre/Mayaro strain is closely related to the progenitor of these Bolivian strains that were isolated between 2002 and 2006.

Research paper thumbnail of Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil

Background: Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs re... more Background: Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs recommended as first-line triple therapy for H.pylori treatment, which results in eradication rates close to 80%, varying regionally, principally due to emergency cases and increases of clarithromycin resistant strains. Nucleotide substitutions at the H. pylori domain V of the 23S rRNA fraction are involved in the macrolide resistance and the A2142G and A2143G mutations are predominant in clinical isolates worldwide including in Brazil. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori A2142G and A2143G rDNA 23S mutations using a molecular approach directly on gastric biopsies of dyspeptic patients consecutively attended at Hospital das Clinicas of Marilia, São Paulo, Brazil.

Research paper thumbnail of A PCR-based strategy for Dombrock screening in Brazilian blood donors reveals a novel allele: the DO*A-WL

Journal of Clinical Laboratory Analysis, 2011

Background: Determination of the molecular basis underlying the antigens in the Dombrock blood gr... more Background: Determination of the molecular basis underlying the antigens in the Dombrock blood group system has shown various rearrangements between the alleles associated with DO Ã A and DO Ã B. Based on this, we employed a PCR-based strategy to screen DO alleles (DO Ã A, DO Ã B, HY Ã 1, HY Ã 2 and JO) in Brazilians. Methods: We tested DNA of 278 Brazilian blood donors by PCR-RFLP on plates of 96 wells to determine the 793A/G (DO Ã A/DO Ã B), 323G/T (HY), 350C/T (JO) and 898C/G (HY Ã 1/HY Ã 2) single nucletide polymorphisms. In order to confirm the results sequence analysis was also performed. Results: When samples of these donors were analyzed, a novel allele combination, the DO Ã A allele (793A and 323G)

Research paper thumbnail of Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil

BMC Gastroenterology, 2012

Background: Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing wo... more Background: Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing worldwide, principally as a result of antibiotic resistant bacterium. Tetracycline is generally included in second line H. pylori eradication regimens. In Brazil, a high level of tetracycline resistance (TetR) is mainly associated with AGA926-928TTC 16 S rDNA nucleotide substitutions. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori 16 S rDNA high level TetR genotype using a molecular approach directly on gastric biopsies of dyspeptic patients attending consecutively at Hospital das Clinicas of Marilia, São Paulo, Brazil. Methods: Gastric biopsy specimens of 68 peptic ulcer disease (PUD) and 327 chronic gastritis (CG) patients with a positive histological diagnosis of H. pylori were investigated for TetR 16 S rDNA genotype through a molecular assay based on amplification of a 16 S rDNA 545 bp fragment by polymerase chain reaction and HinfI restriction fragment length polymorphism (PCR/RFLP). Through this assay, AGA926-928TTC 16 S rDNA TetR genotype resulted in a three DNA fragment restriction pattern (281, 227 and 37 bp) and its absence originated two DNA fragments (264 and 281 bp) due to a 16 S rDNA conserved Hinf I restriction site. Results: The 545 bp 16 S rDNA PCR fragment was amplified from 90% of gastric biopsies from histological H. pylori positive patients. HinfI RFLP revealed absence of the AGA926-928TTC H. pylori genotype and PCR products of two patients showed absence of the conserved 16 S rDNA HinfI restriction site. BLASTN sequence analysis of four amplicons (two conserved and two with an unpredicted HinfI restriction pattern) revealed a 99% homology to H. pylori 16 S rDNA from African, North and South American bacterial isolates. A nucleotide substitution abolished the conserved HinfI restriction site in the two PCR fragments with unpredicted HinfI RFLP, resulting in an EcoRI restriction site. Conclusions: H. pylori AGA926-928TTC 16 S rDNA gene substitutions were not found in our population. More research is required to investigate if H. pylori TetR has a different genetic background in our region and if the nucleotide substitutions of the uncultured H. pylori 16 S rRNA partial sequences have biological significance.

Research paper thumbnail of Different risk factors influence peptic ulcer disease development in a Brazilian population

World Journal of Gastroenterology, 2012

Author contributions: Suzuki RB contributed to the collection and processing of biopsy samples, e... more Author contributions: Suzuki RB contributed to the collection and processing of biopsy samples, epidemiological and molecular comparative analyses; Cola RF and Cola LTB contributed with analytic tools and molecular analysis; Ferrari CG contributed with molecular analysis; Ellinger F, Therezo AL and Silva LC contributed equally to the histopathological analysis; Eterovic A contributed with statistical analysis; and Sperança MA designed the research, wrote the paper and contributed to all comparative analyses.

Research paper thumbnail of A highly sensitive Leishmania infantum chagasi isolation method from bone marrow and peripheral blood of adults and children

Journal of infection in developing countries, Jan 24, 2016

Research paper thumbnail of Novel Single Hematophagous Insect RNA Detection Method Supports Its Use as Sentinels to Survey Flaviviruses Circulation

Dengue Fever [Working Title], Apr 10, 2020

Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, con... more Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, contribute to emergence and reemergence of arboviruses worldwide, where Flavivirus is the most prevalent, and its continuous monitoring can help in preventive control strategies. Thus, the aim of this study was to detect flavivirus RNA in single hematophagous insects, which are used as sentinels. Total RNA was extracted from six Aedes aegypti stored since 2003 and from 100 Culicidae and collected through CDC trap in a public park of a Brazilian Northwest city of São Paulo State. Flavivirus was detected through RT/PCR targeting 230-250 bp of the RNA polymerase coding sequence (NS5). PCR amplicons were sequenced by Sanger method, used in comparative analysis over Basic Local Alignment Search Tool (BLAST) in GenBank, and subjected to Neighbor-Joining phylogenetic analyses. Efficiency of Flavivirus diagnosis was confirmed by detection of Dengue virus serotype 2 in Ae. aegypti. From the 100 collected insects, 19 were positive for Culex flavivirus (CxFV). NS5 partial sequence phylogenetic analysis clustered all CxFV in one branch separated from vertebrate flaviviruses, being applicable to the identification of Flavivirus species. The dipteran RNA extraction methodology described in this work supports detection of flaviviruses in single insects maintained in 80% ethanol, which can be used to constant arbovirus surveillance.

Research paper thumbnail of A PCR-based strategy for Dombrock screening in Brazilian blood donors reveals a novel allele: the DO*A-WL

Journal of Clinical Laboratory Analysis, 2011

Background: Determination of the molecular basis underlying the antigens in the Dombrock blood gr... more Background: Determination of the molecular basis underlying the antigens in the Dombrock blood group system has shown various rearrangements between the alleles associated with DO Ã A and DO Ã B. Based on this, we employed a PCR-based strategy to screen DO alleles (DO Ã A, DO Ã B, HY Ã 1, HY Ã 2 and JO) in Brazilians. Methods: We tested DNA of 278 Brazilian blood donors by PCR-RFLP on plates of 96 wells to determine the 793A/G (DO Ã A/DO Ã B), 323G/T (HY), 350C/T (JO) and 898C/G (HY Ã 1/HY Ã 2) single nucletide polymorphisms. In order to confirm the results sequence analysis was also performed. Results: When samples of these donors were analyzed, a novel allele combination, the DO Ã A allele (793A and 323G)

Research paper thumbnail of A Highly Sensitive and Specific Conventional Molecular Diagnosis for Leishmania infantum Chagasi Based on a Single Copy Gene

Journal of Molecular Biomarkers & Diagnosis, 2016

Research paper thumbnail of Nucleic Acid-based Diagnosis and Epidemiology of Infectious Diseases

Nucleic Acids - From Basic Aspects to Laboratory Tools, 2016

Research paper thumbnail of HNA-3 gene frequencies in Brazilians and a new polymerase chain reaction-restriction fragment length polymorphism method for HNA-3a/3b genotyping

Transfusion, 2013

BACKGROUND: HNA-3 antigens are the result of a rs2288904 single-nucleotide polymorphism (SNP) in ... more BACKGROUND: HNA-3 antigens are the result of a rs2288904 single-nucleotide polymorphism (SNP) in the CTL2, and the HNA-3a and HNA-3b variants are encoded by a guanine and adenine at Nucleotide Position 461. Anti-HNA-3 are involved in severe transfusionrelated acute lung injury reactions and in neonatal alloimmune neutropenia. Since the distribution of the HNA-3 system was unknown in South Americans, in this study we determined the frequency of the HNA-3 alleles in Brazilians. STUDY DESIGN AND METHODS: DNA of 500 blood donors, 120 Xikrin Amerindians, 74 Japanese individuals, and 124 African Brazilians were genotyped for rs2288904 by a polymerase chain reaction (PCR)restriction fragment length polymorphism assay. The PCR product was digested with enzyme Taq α 1, specific to nucleotide guanine (HNA-3a).

Research paper thumbnail of Dombrock genotyping in Brazilian blood donors reveals different regional frequencies of the HY allele

Revista Brasileira de Hematologia e Hemoterapia, 2013

The frequencies of the DO alleles in Minas Gerais, a southeastern state of Brazil, were determine... more The frequencies of the DO alleles in Minas Gerais, a southeastern state of Brazil, were determined from the genotyping of 270 blood donors. Genotyping involved polymerase chain reaction and restriction fragment length polymorphism analysis to identify the 323G>T, 350C>T, 793A>G, and 898C>G mutations, which are related to the HY, JO, DO*A/DO*B, and DO*A-WL/DO*B-WL alleles, respectively. Moreover, the frequencies of rare HY and JO alleles were statistically compared using the chi-square test with data from another Brazilian region.

Research paper thumbnail of Rural Triatoma rubrovaria from southern Brazil harbors Trypanosoma cruzi of lineage IIc

The American journal of tropical medicine and hygiene, 2008

Triatoma infestans, the main vector of Chagas disease, has nearly been eliminated from Brazil. Ne... more Triatoma infestans, the main vector of Chagas disease, has nearly been eliminated from Brazil. Nevertheless, other triatominae species are involved in the domiciliation process, including Triatoma rubrovaria in Rio Grande do Sul State (RS). Previous studies showed that 1.6% of the T. rubrovaria specimens collected at the rural district of Quaraí, RS, were naturally infected by Trypanosoma cruzi. In this study, five T. cruzi isolates obtained from infected triatomines were characterized molecularly and biologically. Genotyping of the T. cruzi isolates showed that they belong to lineage IIc of T. cruzi (TCIIc). Biological characterization showed miotropism and myositis during acute and chronic phases of infection, respectively. Virulence and mortality rates were variable among isolates. To our knowledge, this study corresponds to the first characterization of T. cruzi isolates from T. rubrovaria and the first description of TCIIc in the sylvatic cycle of T. cruzi from the southern reg...

Research paper thumbnail of Isolation and Characterization of Mayaro Virus from a Human in Acre, Brazil

The American journal of tropical medicine and hygiene, Jan 15, 2014

Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of ... more Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of Mayaro fever, an acute febrile illness often presenting with arthralgic manifestations. The true incidence of MAYV infection is likely grossly underestimated because the symptomatic presentation is very similar to that of dengue fever and other acute febrile tropical diseases. We report the complete genome sequence of a MAYV isolate detected from an Acrelândia patient presenting with fever, chills, and sweating, but with no arthralgia. Results show that this isolate belongs to genotype D and is closely related to Bolivian strains. Our results suggest that the Acre/Mayaro strain is closely related to the progenitor of these Bolivian strains that were isolated between 2002 and 2006.

Research paper thumbnail of Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil

Background: Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs re... more Background: Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs recommended as first-line triple therapy for H.pylori treatment, which results in eradication rates close to 80%, varying regionally, principally due to emergency cases and increases of clarithromycin resistant strains. Nucleotide substitutions at the H. pylori domain V of the 23S rRNA fraction are involved in the macrolide resistance and the A2142G and A2143G mutations are predominant in clinical isolates worldwide including in Brazil. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori A2142G and A2143G rDNA 23S mutations using a molecular approach directly on gastric biopsies of dyspeptic patients consecutively attended at Hospital das Clinicas of Marilia, São Paulo, Brazil.

Research paper thumbnail of A PCR-based strategy for Dombrock screening in Brazilian blood donors reveals a novel allele: the DO*A-WL

Journal of Clinical Laboratory Analysis, 2011

Background: Determination of the molecular basis underlying the antigens in the Dombrock blood gr... more Background: Determination of the molecular basis underlying the antigens in the Dombrock blood group system has shown various rearrangements between the alleles associated with DO Ã A and DO Ã B. Based on this, we employed a PCR-based strategy to screen DO alleles (DO Ã A, DO Ã B, HY Ã 1, HY Ã 2 and JO) in Brazilians. Methods: We tested DNA of 278 Brazilian blood donors by PCR-RFLP on plates of 96 wells to determine the 793A/G (DO Ã A/DO Ã B), 323G/T (HY), 350C/T (JO) and 898C/G (HY Ã 1/HY Ã 2) single nucletide polymorphisms. In order to confirm the results sequence analysis was also performed. Results: When samples of these donors were analyzed, a novel allele combination, the DO Ã A allele (793A and 323G)

Research paper thumbnail of Absence of Helicobacter pylori high tetracycline resistant 16S rDNA AGA926-928TTC genotype in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil

BMC Gastroenterology, 2012

Background: Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing wo... more Background: Treatment effectiveness of Helicobacter pylori varies regionally and is decreasing worldwide, principally as a result of antibiotic resistant bacterium. Tetracycline is generally included in second line H. pylori eradication regimens. In Brazil, a high level of tetracycline resistance (TetR) is mainly associated with AGA926-928TTC 16 S rDNA nucleotide substitutions. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori 16 S rDNA high level TetR genotype using a molecular approach directly on gastric biopsies of dyspeptic patients attending consecutively at Hospital das Clinicas of Marilia, São Paulo, Brazil. Methods: Gastric biopsy specimens of 68 peptic ulcer disease (PUD) and 327 chronic gastritis (CG) patients with a positive histological diagnosis of H. pylori were investigated for TetR 16 S rDNA genotype through a molecular assay based on amplification of a 16 S rDNA 545 bp fragment by polymerase chain reaction and HinfI restriction fragment length polymorphism (PCR/RFLP). Through this assay, AGA926-928TTC 16 S rDNA TetR genotype resulted in a three DNA fragment restriction pattern (281, 227 and 37 bp) and its absence originated two DNA fragments (264 and 281 bp) due to a 16 S rDNA conserved Hinf I restriction site. Results: The 545 bp 16 S rDNA PCR fragment was amplified from 90% of gastric biopsies from histological H. pylori positive patients. HinfI RFLP revealed absence of the AGA926-928TTC H. pylori genotype and PCR products of two patients showed absence of the conserved 16 S rDNA HinfI restriction site. BLASTN sequence analysis of four amplicons (two conserved and two with an unpredicted HinfI restriction pattern) revealed a 99% homology to H. pylori 16 S rDNA from African, North and South American bacterial isolates. A nucleotide substitution abolished the conserved HinfI restriction site in the two PCR fragments with unpredicted HinfI RFLP, resulting in an EcoRI restriction site. Conclusions: H. pylori AGA926-928TTC 16 S rDNA gene substitutions were not found in our population. More research is required to investigate if H. pylori TetR has a different genetic background in our region and if the nucleotide substitutions of the uncultured H. pylori 16 S rRNA partial sequences have biological significance.

Research paper thumbnail of Different risk factors influence peptic ulcer disease development in a Brazilian population

World Journal of Gastroenterology, 2012

Author contributions: Suzuki RB contributed to the collection and processing of biopsy samples, e... more Author contributions: Suzuki RB contributed to the collection and processing of biopsy samples, epidemiological and molecular comparative analyses; Cola RF and Cola LTB contributed with analytic tools and molecular analysis; Ferrari CG contributed with molecular analysis; Ellinger F, Therezo AL and Silva LC contributed equally to the histopathological analysis; Eterovic A contributed with statistical analysis; and Sperança MA designed the research, wrote the paper and contributed to all comparative analyses.