The PneuCarriage Project : A multi-centre comparative study to identify the best serotyping methods for examining pneumococcal carriage in vaccine evaluation studies
- Satzke, Catherine, Dunne, Eileen, Porter, Barbara, Klugman, Keith, Mulholland, Kim, PneuCarriage project group, Greenhill, Andrew
- Authors: Satzke, Catherine , Dunne, Eileen , Porter, Barbara , Klugman, Keith , Mulholland, Kim , PneuCarriage project group , Greenhill, Andrew
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS Medicine Vol. 12, no. 11 (2015), p. 1-30
- Full Text:
- Reviewed:
- Description: Background: The pneumococcus is a diverse pathogen whose primary niche is the nasopharynx. Over 90 different serotypes exist, and nasopharyngeal carriage of multiple serotypes is common. Understanding pneumococcal carriage is essential for evaluating the impact of pneumococcal vaccines. Traditional serotyping methods are cumbersome and insufficient for detecting multiple serotype carriage, and there are few data comparing the new methods that have been developed over the past decade. We established the PneuCarriage project, a large, international multi-centre study dedicated to the identification of the best pneumococcal serotyping methods for carriage studies. Methods and Findings: Reference sample sets were distributed to 15 research groups for blinded testing. Twenty pneumococcal serotyping methods were used to test 81 laboratory-prepared (spiked) samples. The five top-performing methods were used to test 260 nasopharyngeal (field) samples collected from children in six high-burden countries. Sensitivity and positive predictive value (PPV) were determined for the test methods and the reference method (traditional serotyping of >100 colonies from each sample). For the alternate serotyping methods, the overall sensitivity ranged from 1% to 99% (reference method 98%), and PPV from 8% to 100% (reference method 100%), when testing the spiked samples. Fifteen methods had ≥70% sensitivity to detect the dominant (major) serotype, whilst only eight methods had ≥70% sensitivity to detect minor serotypes. For the field samples, the overall sensitivity ranged from 74.2% to 95.8% (reference method 93.8%), and PPV from 82.2% to 96.4% (reference method 99.6%). The microarray had the highest sensitivity (95.8%) and high PPV (93.7%). The major limitation of this study is that not all of the available alternative serotyping methods were included. Conclusions: Most methods were able to detect the dominant serotype in a sample, but many performed poorly in detecting the minor serotype populations. Microarray with a culture amplification step was the top-performing method. Results from this comprehensive evaluation will inform future vaccine evaluation and impact studies, particularly in low-income settings, where pneumococcal disease burden remains high. © 2015 Satzke et al. *For a complete list of authors, please see acknowledgments in the published article.
- Authors: Satzke, Catherine , Dunne, Eileen , Porter, Barbara , Klugman, Keith , Mulholland, Kim , PneuCarriage project group , Greenhill, Andrew
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS Medicine Vol. 12, no. 11 (2015), p. 1-30
- Full Text:
- Reviewed:
- Description: Background: The pneumococcus is a diverse pathogen whose primary niche is the nasopharynx. Over 90 different serotypes exist, and nasopharyngeal carriage of multiple serotypes is common. Understanding pneumococcal carriage is essential for evaluating the impact of pneumococcal vaccines. Traditional serotyping methods are cumbersome and insufficient for detecting multiple serotype carriage, and there are few data comparing the new methods that have been developed over the past decade. We established the PneuCarriage project, a large, international multi-centre study dedicated to the identification of the best pneumococcal serotyping methods for carriage studies. Methods and Findings: Reference sample sets were distributed to 15 research groups for blinded testing. Twenty pneumococcal serotyping methods were used to test 81 laboratory-prepared (spiked) samples. The five top-performing methods were used to test 260 nasopharyngeal (field) samples collected from children in six high-burden countries. Sensitivity and positive predictive value (PPV) were determined for the test methods and the reference method (traditional serotyping of >100 colonies from each sample). For the alternate serotyping methods, the overall sensitivity ranged from 1% to 99% (reference method 98%), and PPV from 8% to 100% (reference method 100%), when testing the spiked samples. Fifteen methods had ≥70% sensitivity to detect the dominant (major) serotype, whilst only eight methods had ≥70% sensitivity to detect minor serotypes. For the field samples, the overall sensitivity ranged from 74.2% to 95.8% (reference method 93.8%), and PPV from 82.2% to 96.4% (reference method 99.6%). The microarray had the highest sensitivity (95.8%) and high PPV (93.7%). The major limitation of this study is that not all of the available alternative serotyping methods were included. Conclusions: Most methods were able to detect the dominant serotype in a sample, but many performed poorly in detecting the minor serotype populations. Microarray with a culture amplification step was the top-performing method. Results from this comprehensive evaluation will inform future vaccine evaluation and impact studies, particularly in low-income settings, where pneumococcal disease burden remains high. © 2015 Satzke et al. *For a complete list of authors, please see acknowledgments in the published article.
Characterization of the gut microbiota of Papua New Guineans using reverse transcription quantitative PCR
- Greenhill, Andrew, Tsuji, Hirokazu, Ogata, Kiyohito, Natsuhara, Kazumi, Morita, Ayako, Soli, Kevin, Larkins, Jo-Ann, Tadokoro, Kiyoshi, Odani, Shingo, Baba, Jun, Naito, Yuichi, Tomitsuka, Eriko, Nomoto, Kriko, Siba, Peter, Horwood, Paul, Umezaki, Masahiro
- Authors: Greenhill, Andrew , Tsuji, Hirokazu , Ogata, Kiyohito , Natsuhara, Kazumi , Morita, Ayako , Soli, Kevin , Larkins, Jo-Ann , Tadokoro, Kiyoshi , Odani, Shingo , Baba, Jun , Naito, Yuichi , Tomitsuka, Eriko , Nomoto, Kriko , Siba, Peter , Horwood, Paul , Umezaki, Masahiro
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS ONE Vol. 10, no. 2 (2015), p. 1-15
- Full Text:
- Reviewed:
- Description: There has been considerable interest in composition of gut microbiota in recent years, leading to a better understanding of the role the gut microbiota plays in health and disease. Most studies have been limited in their geographical and socioeconomic diversity to high-income settings, and have been conducted using small sample sizes. To date, few analyses have been conducted in low-income settings, where a better understanding of the gut microbiome could lead to the greatest return in terms of health benefits. Here, we have used quantitative real-time polymerase chain reaction targeting dominant and sub-dominant groups of microorganisms associated with human gut microbiome in 115 people living a subsistence lifestyle in rural areas of Papua New Guinea. Quantification of Clostridium coccoides group, C. leptum subgroup, C. perfringens, Bacteroides fragilis group, Bifidobacterium, Atopobium cluster, Prevotella, Enterobacteriaceae, Enterococcus, Staphylococcus, and Lactobacillus spp. was conducted. Principle coordinates analysis (PCoA) revealed two dimensions with Prevotella, clostridia, Atopobium, Enterobacteriaceae, Enterococcus and Staphylococcus grouping in one dimension, while B. fragilis, Bifidobacterium and Lactobacillus grouping in the second dimension. Highland people had higher numbers of most groups of bacteria detected, and this is likely a key factor for the differences revealed by PCoA between highland and lowland study participants. Age and sex were not major determinants in microbial population composition. The study demonstrates a gut microbial composition with some similarities to those observed in other low-income settings where traditional diets are consumed, which have previously been suggested to favor energy extraction from a carbohydrate rich diet. © 2015 PLOS ONE.
- Authors: Greenhill, Andrew , Tsuji, Hirokazu , Ogata, Kiyohito , Natsuhara, Kazumi , Morita, Ayako , Soli, Kevin , Larkins, Jo-Ann , Tadokoro, Kiyoshi , Odani, Shingo , Baba, Jun , Naito, Yuichi , Tomitsuka, Eriko , Nomoto, Kriko , Siba, Peter , Horwood, Paul , Umezaki, Masahiro
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS ONE Vol. 10, no. 2 (2015), p. 1-15
- Full Text:
- Reviewed:
- Description: There has been considerable interest in composition of gut microbiota in recent years, leading to a better understanding of the role the gut microbiota plays in health and disease. Most studies have been limited in their geographical and socioeconomic diversity to high-income settings, and have been conducted using small sample sizes. To date, few analyses have been conducted in low-income settings, where a better understanding of the gut microbiome could lead to the greatest return in terms of health benefits. Here, we have used quantitative real-time polymerase chain reaction targeting dominant and sub-dominant groups of microorganisms associated with human gut microbiome in 115 people living a subsistence lifestyle in rural areas of Papua New Guinea. Quantification of Clostridium coccoides group, C. leptum subgroup, C. perfringens, Bacteroides fragilis group, Bifidobacterium, Atopobium cluster, Prevotella, Enterobacteriaceae, Enterococcus, Staphylococcus, and Lactobacillus spp. was conducted. Principle coordinates analysis (PCoA) revealed two dimensions with Prevotella, clostridia, Atopobium, Enterobacteriaceae, Enterococcus and Staphylococcus grouping in one dimension, while B. fragilis, Bifidobacterium and Lactobacillus grouping in the second dimension. Highland people had higher numbers of most groups of bacteria detected, and this is likely a key factor for the differences revealed by PCoA between highland and lowland study participants. Age and sex were not major determinants in microbial population composition. The study demonstrates a gut microbial composition with some similarities to those observed in other low-income settings where traditional diets are consumed, which have previously been suggested to favor energy extraction from a carbohydrate rich diet. © 2015 PLOS ONE.
Whole genome sequence analysis of Salmonella Typhi in Papua New Guinea reveals an established population of genotype 2.1.7 sensitive to antimicrobials
- Dyson, Zoe, Malau, Elisheba, Horwood, Paul, Ford, Rebecca, Siba, Valentine, Yoannes, Mition, Pomat, William, Passey, Megan, Judd, Louise, Ingle, Danielle, Williamson, Deborah, Dougan, Gordon, Greenhill, Andrew, Holt, Kathryn
- Authors: Dyson, Zoe , Malau, Elisheba , Horwood, Paul , Ford, Rebecca , Siba, Valentine , Yoannes, Mition , Pomat, William , Passey, Megan , Judd, Louise , Ingle, Danielle , Williamson, Deborah , Dougan, Gordon , Greenhill, Andrew , Holt, Kathryn
- Date: 2022
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases Vol. 16, no. 3 (2022), p.
- Full Text:
- Reviewed:
- Description: Background Typhoid fever, a systemic infection caused by Salmonella enterica serovar Typhi, remains a considerable public health threat in impoverished regions within many low-and middle-income settings. However, we still lack a detailed understanding of the emergence, population structure, molecular mechanisms of antimicrobial resistance (AMR), and transmission dynamics of S. Typhi across many settings, particularly throughout the Asia-Pacific islands. Here we present a comprehensive whole genome sequence (WGS) based overview of S. Typhi populations circulating in Papua New Guinea (PNG) over 30 years. Principle findings Bioinformatic analysis of 86 S. Typhi isolates collected between 1980–2010 demonstrated that the population structure of PNG is dominated by a single genotype (2.1.7) that appears to have emerged in the Indonesian archipelago in the mid-twentieth century with minimal evidence of inter-country transmission. Genotypic and phenotypic data demonstrated that the PNG S. Typhi population appears to be susceptible to former first line drugs for treating typhoid fever (chloramphenicol, ampicillin and co-trimoxazole), as well as fluoroquinolones, third generation cephalosporins, and macrolides. PNG genotype 2.1.7 was genetically con-served, with very few deletions, and no evidence of plasmid or prophage acquisition. Genetic variation among this population was attributed to either single point mutations, or homologous recombination adjacent to repetitive ribosomal RNA operons. Significance Antimicrobials remain an effective option for the treatment of typhoid fever in PNG, along with other intervention strategies including improvements to water, sanitation and hygiene (WaSH) related infrastructure and potentially the introduction of Vi-conjugate vaccines. However, continued genomic surveillance is warranted to monitor for the emergence of AMR within local populations, or the introduction of AMR associated genotypes of S. Typhi in this setting. © The Authors.
- Authors: Dyson, Zoe , Malau, Elisheba , Horwood, Paul , Ford, Rebecca , Siba, Valentine , Yoannes, Mition , Pomat, William , Passey, Megan , Judd, Louise , Ingle, Danielle , Williamson, Deborah , Dougan, Gordon , Greenhill, Andrew , Holt, Kathryn
- Date: 2022
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases Vol. 16, no. 3 (2022), p.
- Full Text:
- Reviewed:
- Description: Background Typhoid fever, a systemic infection caused by Salmonella enterica serovar Typhi, remains a considerable public health threat in impoverished regions within many low-and middle-income settings. However, we still lack a detailed understanding of the emergence, population structure, molecular mechanisms of antimicrobial resistance (AMR), and transmission dynamics of S. Typhi across many settings, particularly throughout the Asia-Pacific islands. Here we present a comprehensive whole genome sequence (WGS) based overview of S. Typhi populations circulating in Papua New Guinea (PNG) over 30 years. Principle findings Bioinformatic analysis of 86 S. Typhi isolates collected between 1980–2010 demonstrated that the population structure of PNG is dominated by a single genotype (2.1.7) that appears to have emerged in the Indonesian archipelago in the mid-twentieth century with minimal evidence of inter-country transmission. Genotypic and phenotypic data demonstrated that the PNG S. Typhi population appears to be susceptible to former first line drugs for treating typhoid fever (chloramphenicol, ampicillin and co-trimoxazole), as well as fluoroquinolones, third generation cephalosporins, and macrolides. PNG genotype 2.1.7 was genetically con-served, with very few deletions, and no evidence of plasmid or prophage acquisition. Genetic variation among this population was attributed to either single point mutations, or homologous recombination adjacent to repetitive ribosomal RNA operons. Significance Antimicrobials remain an effective option for the treatment of typhoid fever in PNG, along with other intervention strategies including improvements to water, sanitation and hygiene (WaSH) related infrastructure and potentially the introduction of Vi-conjugate vaccines. However, continued genomic surveillance is warranted to monitor for the emergence of AMR within local populations, or the introduction of AMR associated genotypes of S. Typhi in this setting. © The Authors.
- «
- ‹
- 1
- ›
- »