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2.
MMWR Recomm Rep ; 73(1): 1-32, 2024 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-38319847

RESUMO

This report provides new CDC recommendations for tests that can support a diagnosis of syphilis, including serologic testing and methods for the identification of the causative agent Treponema pallidum. These comprehensive recommendations are the first published by CDC on laboratory testing for syphilis, which has traditionally been based on serologic algorithms to detect a humoral immune response to T. pallidum. These tests can be divided into nontreponemal and treponemal tests depending on whether they detect antibodies that are broadly reactive to lipoidal antigens shared by both host and T. pallidum or antibodies specific to T. pallidum, respectively. Both types of tests must be used in conjunction to help distinguish between an untreated infection or a past infection that has been successfully treated. Newer serologic tests allow for laboratory automation but must be used in an algorithm, which also can involve older manual serologic tests. Direct detection of T. pallidum continues to evolve from microscopic examination of material from lesions for visualization of T. pallidum to molecular detection of the organism. Limited point-of-care tests for syphilis are available in the United States; increased availability of point-of-care tests that are sensitive and specific could facilitate expansion of screening programs and reduce the time from test result to treatment. These recommendations are intended for use by clinical laboratory directors, laboratory staff, clinicians, and disease control personnel who must choose among the multiple available testing methods, establish standard operating procedures for collecting and processing specimens, interpret test results for laboratory reporting, and counsel and treat patients. Future revisions to these recommendations will be based on new research or technologic advancements for syphilis clinical laboratory science.


Assuntos
Sífilis , Humanos , Estados Unidos , Sífilis/diagnóstico , Sorodiagnóstico da Sífilis/métodos , Treponema pallidum , Testes Sorológicos , Centers for Disease Control and Prevention, U.S.
3.
Sci Rep ; 14(1): 3702, 2024 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-38355640

RESUMO

The transmission risk of SARS-CoV-2 within hospitals can exceed that in the general community because of more frequent close proximity interactions (CPIs). However, epidemic risk across wards is still poorly described. We measured CPIs directly using wearable sensors given to all present in a clinical ward over a 36-h period, across 15 wards in three hospitals in April-June 2020. Data were collected from 2114 participants and combined with a simple transmission model describing the arrival of a single index case to the ward to estimate the risk of an outbreak. Estimated epidemic risk ranged four-fold, from 0.12 secondary infections per day in an adult emergency to 0.49 per day in general paediatrics. The risk presented by an index case in a patient varied 20-fold across wards. Using simulation, we assessed the potential impact on outbreak risk of targeting the most connected individuals for prevention. We found that targeting those with the highest cumulative contact hours was most impactful (20% reduction for 5% of the population targeted), and on average resources were better spent targeting patients. This study reveals patterns of interactions between individuals in hospital during a pandemic and opens new routes for research into airborne nosocomial risk.


Assuntos
Hospitais , SARS-CoV-2 , Adulto , Humanos , Criança , Surtos de Doenças , Pandemias/prevenção & controle
5.
PLoS Genet ; 20(1): e1011125, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38236819

RESUMO

[This corrects the article DOI: 10.1371/journal.pgen.1010751.].

7.
New Phytol ; 240(5): 2072-2084, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37793435

RESUMO

Lateral gene transfer (LGT) is the movement of DNA between organisms without sexual reproduction. The acquired genes represent genetic novelties that have independently evolved in the donor's genome. Phylogenetic methods have shown that LGT is widespread across the entire grass family, although we know little about the underlying dynamics. We identify laterally acquired genes in five de novo reference genomes from the same grass genus (four Alloteropsis semialata and one Alloteropsis angusta). Using additional resequencing data for a further 40 Alloteropsis individuals, we place the acquisition of each gene onto a phylogeny using stochastic character mapping, and then infer rates of gains and losses. We detect 168 laterally acquired genes in the five reference genomes (32-100 per genome). Exponential decay models indicate that the rate of LGT acquisitions (6-28 per Ma) and subsequent losses (11-24% per Ma) varied significantly among lineages. Laterally acquired genes were lost at a higher rate than vertically inherited loci (0.02-0.8% per Ma). This high turnover creates intraspecific gene content variation, with a preponderance of them occurring as accessory genes in the Alloteropsis pangenome. This rapid turnover generates standing variation that can ultimately fuel local adaptation.


Assuntos
Transferência Genética Horizontal , Poaceae , Humanos , Filogenia , Poaceae/genética , Genoma , Evolução Molecular
9.
Ann Bot ; 132(3): 365-382, 2023 11 23.
Artigo em Inglês | MEDLINE | ID: mdl-37422712

RESUMO

BACKGROUND: Numerous groups of plants have adapted to CO2 limitations by independently evolving C4 photosynthesis. This trait relies on concerted changes in anatomy and biochemistry to concentrate CO2 within the leaf and thereby boost productivity in tropical conditions. The ecological and economic importance of C4 photosynthesis has motivated intense research, often relying on comparisons between distantly related C4 and non-C4 plants. The photosynthetic type is fixed in most species, with the notable exception of the grass Alloteropsis semialata. This species includes populations exhibiting the ancestral C3 state in southern Africa, intermediate populations in the Zambezian region and C4 populations spread around the palaeotropics. SCOPE: We compile here the knowledge on the distribution and evolutionary history of the Alloteropsis genus as a whole and discuss how this has furthered our understanding of C4 evolution. We then present a chromosome-level reference genome for a C3 individual and compare the genomic architecture with that of a C4 accession of A. semialata. CONCLUSIONS: Alloteropsis semialata is one of the best systems in which to investigate the evolution of C4 photosynthesis because the genetic and phenotypic variation provides a fertile ground for comparative and population-level studies. Preliminary comparative genomic investigations show that the C3 and C4 genomes are highly syntenic and have undergone a modest amount of gene duplication and translocation since the different photosynthetic groups diverged. The background knowledge and publicly available genomic resources make A. semialata a great model for further comparative analyses of photosynthetic diversification.


Assuntos
Dióxido de Carbono , Poaceae , Poaceae/genética , Plantas , Fotossíntese/genética , Fenótipo
10.
PLoS Genet ; 19(5): e1010751, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-37141297

RESUMO

Methyl salicylate is an important inter- and intra-plant signaling molecule, but is deemed undesirable by humans when it accumulates to high levels in ripe fruits. Balancing the tradeoff between consumer satisfaction and overall plant health is challenging as the mechanisms regulating volatile levels have not yet been fully elucidated. In this study, we investigated the accumulation of methyl salicylate in ripe fruits of tomatoes that belong to the red-fruited clade. We determine the genetic diversity and the interaction of four known loci controlling methyl salicylate levels in ripe fruits. In addition to Non-Smoky Glucosyl Transferase 1 (NSGT1), we uncovered extensive genome structural variation (SV) at the Methylesterase (MES) locus. This locus contains four tandemly duplicated Methylesterase genes and genome sequence investigations at the locus identified nine distinct haplotypes. Based on gene expression and results from biparental crosses, functional and non-functional haplotypes for MES were identified. The combination of the non-functional MES haplotype 2 and the non-functional NSGT1 haplotype IV or V in a GWAS panel showed high methyl salicylate levels in ripe fruits, particularly in accessions from Ecuador, demonstrating a strong interaction between these two loci and suggesting an ecological advantage. The genetic variation at the other two known loci, Salicylic Acid Methyl Transferase 1 (SAMT1) and tomato UDP Glycosyl Transferase 5 (SlUGT5), did not explain volatile variation in the red-fruited tomato germplasm, suggesting a minor role in methyl salicylate production in red-fruited tomato. Lastly, we found that most heirloom and modern tomato accessions carried a functional MES and a non-functional NSGT1 haplotype, ensuring acceptable levels of methyl salicylate in fruits. Yet, future selection of the functional NSGT1 allele could potentially improve flavor in the modern germplasm.


Assuntos
Solanum lycopersicum , Humanos , Solanum lycopersicum/genética , Salicilatos/análise , Salicilatos/química , Salicilatos/metabolismo , Glicosiltransferases , Equador , Frutas/genética
11.
Plant Cell Environ ; 46(8): 2310-2322, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37184423

RESUMO

C4 photosynthesis results from anatomical and biochemical characteristics that together concentrate CO2 around ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), increasing productivity in warm conditions. This complex trait evolved through the gradual accumulation of components, and particular species possess only some of these, resulting in weak C4 activity. The consequences of adding C4 components have been modelled and investigated through comparative approaches, but the intraspecific dynamics responsible for strengthening the C4 pathway remain largely unexplored. Here, we evaluate the link between anatomical variation and C4 activity, focusing on populations of the photosynthetically diverse grass Alloteropsis semialata that fix various proportions of carbon via the C4 cycle. The carbon isotope ratios in these populations range from values typical of C3 to those typical of C4 plants. This variation is statistically explained by a combination of leaf anatomical traits linked to the preponderance of bundle sheath tissue. We hypothesize that increased investment in bundle sheath boosts the strength of the intercellular C4 pump and shifts the balance of carbon acquisition towards the C4 cycle. Carbon isotope ratios indicating a stronger C4 pathway are associated with warmer, drier environments, suggesting that incremental anatomical alterations can lead to the emergence of C4 physiology during local adaptation within metapopulations.


Assuntos
Plantas , Poaceae , Poaceae/metabolismo , Plantas/metabolismo , Fotossíntese/fisiologia , Carbono/metabolismo , Isótopos de Carbono/metabolismo , Folhas de Planta/metabolismo , Dióxido de Carbono/metabolismo
12.
J Clin Microbiol ; 61(6): e0016823, 2023 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-37219422

RESUMO

Automated nontreponemal rapid plasma reagin (RPR) tests were recently introduced in the United States for syphilis testing and limited performance data are available. In collaboration with the Association of Public Health Laboratories, three public health laboratories (PHL) were chosen through a competitive selection process to evaluate the performance of three FDA-cleared automated RPR test systems: BioPlex 2200 Syphilis Total & RPR assay (Bio-Rad Laboratories), AIX 1000 (Gold Standard Diagnostics), and ASI Evolution (Arlington Scientific). Panels prepared at the CDC included: a qualitative panel comprised of 734 syphilis reactive/nonreactive sera; a quantitative panel of 50 syphilis reactive sera (RPR titer 1:64 to 1:1,024); and a reproducibility panel of 15 nonreactive and reactive sera (RPR titer 1:1 to 1:64). Panels were shipped frozen to the PHL and tested on the automated RPR systems following manufacturers' instructions. Prior test results were blinded to all laboratories. When compared to manual RPR (Arlington Scientific) performed at the CDC as a reference test, the qualitative panel results demonstrated an overall concordance of 95.9% for AIX 1000, 94.6% for ASI Evolution, and 92.6% for Bioplex RPR; quantitative panel showed within range titer of 2-fold for 94% of specimens for AIX 1000, 68% for ASI Evolution, and 64% for BioPlex RPR, and the reproducibility testing panel demonstrated point estimates ranging from 69 to 95%. Automated RPR instruments could reduce turnaround time and minimize interpretation errors. However, additional evaluations with more specimens could assist laboratories with implementing automated RPR tests and understanding their limitations.


Assuntos
Sífilis , Humanos , Sífilis/diagnóstico , Reaginas , Reprodutibilidade dos Testes , Anticorpos Antibacterianos , Sorodiagnóstico da Sífilis/métodos , Treponema pallidum
13.
15.
Online braz. j. nurs. (Online) ; 22: e20236641, 01 jan 2023. ilus
Artigo em Inglês, Português | LILACS, BDENF - enfermagem (Brasil) | ID: biblio-1437872

RESUMO

OBJETIVO: Mapear as competências do enfermeiro especialista em Dermatologia. MÉTODO: Trata-se de uma scoping review, desenvolvida conforme as recomendações do Joanna Briggs Institute (JBI). Para elaboração da questão de pesquisa utilizou-se a estratégia Population, Concept e Context. A busca por publicações, sem limitar idioma e recorte temporal, foi realizada nas fontes: The Cumulative Index to Nursing and Allied Health Literature (CINAHL), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), National Library of Medicine (MEDLINE) via PubMed, Scopus, Embase e Cochrane Library, além de sites das associações de especialistas e no Google Acadêmico. RESULTADOS: Foram selecionadas 17 publicações, originando as categorias: avaliação integral do paciente dermatológico, prescrição de medicamentos, detecção precoce e prevenção do câncer de pele, tratamento de feridas e estética. CONCLUSÃO: Diante da vasta possibilidade de atuação do enfermeiro nesta área, constata-se a necessidade de ampliação da discussão e de pesquisas sobre o tema.


OBJECTIVE: To map the competencies of specialist nurses in Dermatology. METHOD: A scoping review was conducted according to the recommendations of the Joanna Briggs Institute (JBI). The Population, Concept, and Context framework established the research question. The search for publications was carried out in different sources, including Cumulative Index to Nursing and Allied Health Literature (CINAHL), Latin America and the Caribbean Literature on Health Sciences (LILACS), National Library of Medicine (MEDLINE) via PubMed, Scopus, Embase, and the Cochrane Library. Additionally, expert association websites and Google Scholar were searched. No language or time restrictions were applied. RESULTS: Seventeen publications were selected, originating the categories: integral evaluation of the dermatological patient, drug prescription, early detection and prevention of skin cancer, wound care, and aesthetics. CONCLUSION: Faced with the vast possibilities for nurses to work in Dermatology, there is a need to expand the discussion and research on the subject.


Assuntos
Competência Profissional , Especialização , Competência Clínica , Atenção à Saúde , Dermatologia , Enfermeiras e Enfermeiros
16.
Plant Physiol ; 191(1): 110-124, 2023 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-36315067

RESUMO

Methyl salicylate imparts a potent flavor and aroma described as medicinal and wintergreen that is undesirable in tomato (Solanum lycopersicum) fruit. Plants control the quantities of methyl salicylate through a variety of biosynthetic pathways, including the methylation of salicylic acid to form methyl salicylate and subsequent glycosylation to prevent methyl salicylate emission. Here, we identified a subclade of tomato methyl esterases, SALICYLIC ACID METHYL ESTERASE1-4, responsible for demethylation of methyl salicylate to form salicylic acid in fruits. This family was identified by proximity to a highly significant methyl salicylate genome-wide association study locus on chromosome 2. Genetic mapping studies in a biparental population confirmed a major methyl salicylate locus on chromosome 2. Fruits from SlMES1 knockout lines emitted significantly (P < 0,05, t test) higher amounts of methyl salicylate than wild-type fruits. Double and triple mutants of SlMES2, SlMES3, and SlMES4 emitted even more methyl salicylate than SlMES1 single knockouts-but not at statistically distinguishable levels-compared to the single mutant. Heterologously expressed SlMES1 and SlMES3 acted on methyl salicylate in vitro, with SlMES1 having a higher affinity for methyl salicylate than SlMES3. The SlMES locus has undergone major rearrangement, as demonstrated by genome structure analysis in the parents of the biparental population. Analysis of accessions that produce high or low levels of methyl salicylate showed that SlMES1 and SlMES3 genes expressed the highest in the low methyl salicylate lines. None of the MES genes were appreciably expressed in the high methyl salicylate-producing lines. We concluded that the SlMES gene family encodes tomato methyl esterases that convert methyl salicylate to salicylic acid in ripe tomato fruit. Their ability to decrease methyl salicylate levels by conversion to salicylic acid is an attractive breeding target to lower the level of a negative contributor to flavor.


Assuntos
Ácido Salicílico , Solanum lycopersicum , Ácido Salicílico/metabolismo , Solanum lycopersicum/genética , Frutas/genética , Frutas/metabolismo , Estudo de Associação Genômica Ampla , Melhoramento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
17.
Plants (Basel) ; 11(22)2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36432848

RESUMO

Melon is an economically important crop with widely diverse fruit morphology and ripening characteristics. Its diploid sequenced genome and multiple genomic tools make this species suitable to study the genetic architecture of fruit traits. With the development of this introgression line population of the elite varieties 'Piel de Sapo' and 'Védrantais', we present a powerful tool to study fruit morphology and ripening traits that can also facilitate characterization or pyramidation of QTLs in inodorous melon types. The population consists of 36 lines covering almost 98% of the melon genome, with an average of three introgressions per chromosome and segregating for multiple fruit traits: morphology, ripening and quality. High variability in fruit morphology was found within the population, with 24 QTLs affecting six different traits, confirming previously reported QTLs and two newly detected QTLs, FLQW5.1 and FWQW7.1. We detected 20 QTLs affecting fruit ripening traits, six of them reported for the first time, two affecting the timing of yellowing of the rind (EYELLQW1.1 and EYELLQW8.1) and four at the end of chromosome 8 affecting aroma, abscission and harvest date (EAROQW8.3, EALFQW8.3, ABSQW8.3 and HARQW8.3). We also confirmed the location of several QTLs, such as fruit-quality-related QTLs affecting rind and flesh appearance and flesh firmness.

18.
PLoS One ; 17(10): e0276894, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36301973

RESUMO

BACKGROUND: A significant increase in distress and mental health illnesses has been identified in medical students during their training. As a result, medical schools have attempted to understand factors linked to well-being. Wellness questionnaires present a useful approach to identifying students with risk factors for mental health to provide appropriate resources for support and referrals. This study aims to identify validated questionnaires in the literature that measure medical student wellness. METHODS: A scoping review methodology was selected and an exhaustive search of MEDLINE, Embase, CINAHL, APA PsycInfo, EPIC, and Education Source, was performed from 1999 to May 27, 2021. A compilation of validated wellness evaluation tools, surveys and questionnaires assessing wellness beyond depression and anxiety was reviewed. All validated methods of wellness assessment for medical students were included. RESULTS: 5,001 studies were identified once duplicate records were removed. After applying inclusion and exclusion criteria, 23 articles were included in a qualitative synthesis and explored in detail. The following six validated questionnaires measuring the wellness of medical school students are reported and discussed: the Medical Student Stress Profile (MSSP), the Medical Student Stress Questionnaire (MSSQ), the Medical Student Well-Being Index (MSWBI), the Perceived Medical School Stress (PMSS), the Perceived Stress Scale for Medical Students (PSSMS), and the Oldenburg Burnout Inventory-Medical Student Version (OLBI-MS). These validated questionnaires provide various aspects to the assessment of wellbeing. CONCLUSIONS: Wellbeing evaluations are reliable in identifying medical students who are at risk for mental health illnesses but must be chosen carefully based on contexts, academic environment and student population. A direct comparison between validated questionnaires for student wellbeing is not possible and individual medical schools must determine the appropriateness and validity of such tools based on population-specific characteristics and demands.


Assuntos
Esgotamento Profissional , Estudantes de Medicina , Humanos , Estudantes de Medicina/psicologia , Depressão/psicologia , Ansiedade/diagnóstico , Transtornos de Ansiedade , Esgotamento Profissional/psicologia , Inquéritos e Questionários , Faculdades de Medicina
19.
Fisioter. Pesqui. (Online) ; 29(3): 230-238, jul.-set. 2022. tab, graf
Artigo em Português | LILACS-Express | LILACS | ID: biblio-1421475

RESUMO

RESUMO O objetivo deste estudo foi avaliar a prevalência, o conhecimento e os fatores associados à incontinência urinária (IU) em mulheres estudantes de um curso de fisioterapia, bem como sua qualidade de vida. Foi aplicada uma ficha de avaliação, o International Consultation on Incontinence Questionnaire - Short Form (ICIQ-SF) e um questionário de conhecimento sobre a musculatura do assoalho pélvico (MAP) e a ocorrência de IU, baseado em estudo tipo conhecimento, atitude e prática (CAP). Das 248 estudantes do curso, 170 (69%) participaram do estudo. Pela ficha de avaliação, 111 (65%) relataram perda urinária. De acordo com o ICIQ-SF, 63 participantes (37%) são incontinentes, sendo que 41 (65%) dessas apresentam IU de esforço, apresentando pequena perda e com frequência de uma vez na semana ou menos, com baixo impacto na sua qualidade de vida. A maioria das participantes que relataram ter dor na relação sexual (59%) e alguns sintomas uroginecológicos e intestinais, como esforço ao urinar (92%), jato interrompido (75%), incômodo na região vaginal (73%), constipação (53%), esforço ao defecar (53%) e esvaziamento intestinal incompleto (70%), apresentou queixa de IU. Todas as que usam protetor (100%) e procuraram atendimento médico (100%) eram incontinentes. O uso de anticoncepcional foi maior em mulheres sem perda urinária (84%). O conhecimento sobre a MAP e a IU foi adquirido gradualmente com o avançar do curso. Esse resultado sugere que as mulheres que não cursam Fisioterapia não têm domínio do assunto, sendo necessário disseminar o conhecimento sobre a IU e a atuação da fisioterapia.


RESUMEN El objetivo de este estudio fue evaluar la prevalencia, el conocimiento y los factores asociados a la incontinencia urinaria (IU) en mujeres estudiantes de Fisioterapia, así como su calidad de vida. Se aplicó un formulario de evaluación, el International Consultation on Incontinence Questionnaire - Short Form (ICIQ-SF) y un cuestionario de conocimiento sobre la musculatura del suelo pélvico (MSP) y la ocurrencia de IU, basado en un estudio de conocimiento, actitud y práctica (CAP). De las 248 estudiantes del curso, 170 (69%) participaron en el estudio. Según el formulario de evaluación, 111 (65%) reportaron pérdida urinaria. Conforme el ICIQ-SF, 63 participantes (37%) son incontinentes, y 41 (65%) de ellas tienen IU de esfuerzo, presentando una pequeña pérdida y con frecuencia de una vez a la semana o menos, con bajo impacto en su calidad de vida. La mayoría de las participantes que informaron sentir dolor en las relaciones sexuales (59%) y algunos síntomas uroginecológicos e intestinales, como esfuerzo al orinar (92%), chorro interrumpido (75%), molestias en la región vaginal (73%), estreñimiento (53%), esfuerzo al defecar (53%) y vaciamiento intestinal incompleto (70%), se quejó de IU. Las participantes que usan protectores (100%) y buscan atención médica (100%) eran incontinentes. El uso de anticonceptivos fue mayor en las mujeres sin pérdida urinaria (84%). El conocimiento sobre la MSP e IU se adquirió de manera gradual a medida que avanzaba el curso. Este resultado apunta que las mujeres que no estudian Fisioterapia no tienen dominio del tema y es necesario difundir conocimientos sobre la IU y la actuación de la fisioterapia.


ABSTRACT This study aimed to evaluate the prevalence, knowledge, factors associated with urinary incontinence (UI), and quality of life in female students enrolled in a physical therapy undergraduate course. Students had to answer a form we developed, the International Questionnaire on Incontinence Consultation - Short Form (ICIQ-SF), and a knowledge questionnaire on pelvic floor musculature (PFM) and the occurrence of UI, based on a previous knowledge, attitude, and practice (KAP) study. Out of 248 students, 170 (69%) participated in our study. According to our evaluation form, 111 women (65%) reported urinary loss at some point in their lives. According to ICIQ-SF criteria, 63 (37%) women were incontinent, 41 (65%) suffered from strain urinary incontinence, showing small urinary losses once per week or less, which slightly impacted their quality of life. Most women who have pain during sexual intercourse (59%) and some urogynecological and intestinal symptom, such as straining (92%), intermittent urination (75%), nausea (73%), constipation (53%), excessive effort (53%), and incomplete outlet (70%) also suffered from UI. All women who used pads (100%) and sought medical care (100%) were incontinent. The use of contraceptive was higher among women without UI (84%). Participants built their knowledge on PFM and UI as the course progressed. These results suggest that women who attend undergraduate courses outside physical therapy have limited knowledge about the subject. We find it necessary to spread knowledge about UI and the importance of physical therapy to prevent and treat UI.

20.
mSphere ; 7(3): e0000922, 2022 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-35491834

RESUMO

Downstream next-generation sequencing (NGS) of the syphilis spirochete Treponema pallidum subspecies pallidum (T. pallidum) is hindered by low bacterial loads and the overwhelming presence of background metagenomic DNA in clinical specimens. In this study, we investigated selective whole-genome amplification (SWGA) utilizing multiple displacement amplification (MDA) in conjunction with custom oligonucleotides with an increased specificity for the T. pallidum genome and the capture and removal of 5'-C-phosphate-G-3' (CpG) methylated host DNA using the NEBNext Microbiome DNA enrichment kit followed by MDA with the REPLI-g single cell kit as enrichment methods to improve the yields of T. pallidum DNA in isolates and lesion specimens from syphilis patients. Sequencing was performed using the Illumina MiSeq v2 500 cycle or NovaSeq 6000 SP platform. These two enrichment methods led to 93 to 98% genome coverage at 5 reads/site in 5 clinical specimens from the United States and rabbit-propagated isolates, containing >14 T. pallidum genomic copies/µL of sample for SWGA and >129 genomic copies/µL for CpG methylation capture with MDA. Variant analysis using sequencing data derived from SWGA-enriched specimens showed that all 5 clinical strains had the A2058G mutation associated with azithromycin resistance. SWGA is a robust method that allows direct whole-genome sequencing (WGS) of specimens containing very low numbers of T. pallidum, which has been challenging until now. IMPORTANCE Syphilis is a sexually transmitted, disseminated acute and chronic infection caused by the bacterial pathogen Treponema pallidum subspecies pallidum. Primary syphilis typically presents as single or multiple mucocutaneous lesions and, if left untreated, can progress through multiple stages with various clinical manifestations. Molecular studies often rely on direct amplification of DNA sequences from clinical specimens; however, this can be impacted by inadequate samples due to disease progression or timing of patients seeking clinical care. While genotyping has provided important data on circulating strains over the past 2 decades, WGS data are needed to better understand strain diversity, perform evolutionary tracing, and monitor antimicrobial resistance markers. The significance of our research is the development of an SWGA DNA enrichment method that expands the range of clinical specimens that can be directly sequenced to include samples with low numbers of T. pallidum.


Assuntos
Sífilis , Treponema pallidum , Animais , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Coelhos , Sífilis/microbiologia , Treponema pallidum/genética , Sequenciamento Completo do Genoma
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