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1.
BMC Microbiol ; 22(1): 23, 2022 01 13.
Artículo en Inglés | MEDLINE | ID: mdl-35026981

RESUMEN

BACKGROUND: Streptococcus agalactiae or Group B Streptococcus (GBS) is an encapsulated gram-positive bacterial pathobiont that commonly colonizes the lower gastrointestinal tract and reproductive tract of human hosts. This bacterium can infect the gravid reproductive tract and cause invasive infections of pregnant patients and neonates. Upon colonizing the reproductive tract, the bacterial cell is presented with numerous nutritional challenges imposed by the host. One strategy employed by the host innate immune system is intoxication of bacterial invaders with certain transition metals such as zinc. METHODOLOGY: Previous work has demonstrated that GBS must employ elegant strategies to circumnavigate zinc stress in order to survive in the vertebrate host. We assessed 30 strains of GBS from diverse isolation sources, capsular serotypes, and sequence types for susceptibility or resistance to zinc intoxication. RESULTS: Invasive strains, such as those isolated from early onset disease manifestations of GBS infection were significantly less susceptible to zinc toxicity than colonizing strains isolated from rectovaginal swabs of pregnant patients. Additionally, capsular type III (cpsIII) strains and the ST-17 and ST-19 strains exhibited the greatest resilience to zinc stress, whereas ST-1 and ST-12 strains as well as those possessing capsular type Ib (cpsIb) were more sensitive to zinc intoxication. Thus, this study demonstrates that the transition metal zinc possesses antimicrobial properties against a wide range of GBS strains, with isolation source, capsular serotype, and sequence type contributing to susceptibility or resistance to zinc stress.


Asunto(s)
Antibacterianos/farmacología , Cloruros/farmacocinética , Serogrupo , Streptococcus agalactiae/efectos de los fármacos , Streptococcus agalactiae/genética , Compuestos de Zinc/farmacocinética , Antibacterianos/metabolismo , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/efectos de los fármacos , Cloruros/metabolismo , Femenino , Humanos , Recién Nacido , Pruebas de Sensibilidad Microbiana , Embarazo , Serotipificación , Infecciones Estreptocócicas/sangre , Infecciones Estreptocócicas/tratamiento farmacológico , Infecciones Estreptocócicas/microbiología , Streptococcus agalactiae/clasificación , Streptococcus agalactiae/crecimiento & desarrollo , Vagina/efectos de los fármacos , Vagina/microbiología , Compuestos de Zinc/metabolismo
2.
Infect Immun ; 89(11): e0037421, 2021 10 15.
Artículo en Inglés | MEDLINE | ID: mdl-34338550

RESUMEN

The Streptococcus pneumoniae capsule is essential for disease pathogenesis, suggesting that even minor genetic changes within the cps locus could potentially have important consequences. Arends et al. (D. W. Arends, W. R. Miellet, J. D. Langereis, T. H. A. Ederveen, et al., Infect Immun 89:e00246-21, 2021, https://doi.org/10.1128/IAI.00246-21) have identified 79 different nonsynonymous single-nucleotide polymorphisms (SNPs) in the cps locus of 338 19A serotype strains and shown significant variations between strains in nucleotide sugar content and capsule shedding. Further work is required to characterize whether any of these changes have important functional consequences on capsule-host interactions.


Asunto(s)
Sitios Genéticos , Polimorfismo de Nucleótido Simple , Polisacáridos Bacterianos/genética , Streptococcus pneumoniae/genética , Cápsulas Bacterianas/clasificación , Humanos
3.
mBio ; 12(3): e0080021, 2021 06 29.
Artículo en Inglés | MEDLINE | ID: mdl-34061603

RESUMEN

Capsular polysaccharides (CPSs) are major virulence factors that decorate the surfaces of many human bacterial pathogens. In their pure form or as glycoconjugate vaccines, CPSs are extensively used in vaccines deployed in clinical practice worldwide. However, our understanding of the structural requirements for interactions between CPSs and antibodies is limited. A longstanding model based on comprehensive observations of antibody repertoires binding to CPSs is that antibodies expressing heavy chain variable gene family 3 (VH3) predominate in these binding interactions in humans and VH3 homologs in mice. Toward understanding this highly conserved interaction, we generated a panel of mouse monoclonal antibodies (MAb) against Streptococcus pneumoniae serotype 3 CPS, determined an X-ray crystal structure of a protective MAb in complex with a hexasaccharide derived from enzymatic hydrolysis of the polysaccharide, and elucidated the structural requirements for this binding interaction. The crystal structure revealed a binding pocket containing aromatic side chains, suggesting the importance of hydrophobicity in the interaction. Through mutational analysis, we determined the amino acids that are critical in carbohydrate binding. Through elucidating the structural and functional properties of a panel of murine MAbs, we offer an explanation for the predominant use of the human VH3 gene family in antibodies against CPSs with implications in knowledge-based vaccine design. IMPORTANCE Infectious diseases caused by pathogenic bacteria are a major threat to human health. Capsular polysaccharides (CPSs) of many pathogenic bacteria have been used as the main components of glycoconjugate vaccines against bacterial diseases in clinical practice worldwide, with various degrees of success. Immunization with a glycoconjugate vaccine elicits T cell help for B cells that produce IgG antibodies to the CPS. Thus, it is important to develop an in-depth understanding of the interactions of carbohydrate epitopes with the antibodies. Structural characterization of the ligand binding of polysaccharide-specific antibodies laid out in this study may have fundamental biological implications for our comprehension of how the humoral immune system recognizes polysaccharide antigens, and in future knowledge-based vaccine design.


Asunto(s)
Anticuerpos Antibacterianos/inmunología , Cápsulas Bacterianas/química , Polisacáridos Bacterianos/inmunología , Polisacáridos Bacterianos/metabolismo , Streptococcus pneumoniae/química , Streptococcus pneumoniae/inmunología , Animales , Anticuerpos Monoclonales , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/inmunología , Cristalización , Femenino , Humanos , Ligandos , Ratones , Ratones Endogámicos BALB C , Modelos Estructurales , Polisacáridos Bacterianos/química , Serogrupo , Streptococcus pneumoniae/clasificación , Streptococcus pneumoniae/patogenicidad , Vacunación
4.
Vet Microbiol ; 258: 109121, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-34020174

RESUMEN

This study compares the utility of a commercially available multiplex q-PCR assay for serotyping A1, A2, and A6 M. haemolytica serotypes with indirect hemagglutination, for determining the relative distribution of M. haemolytica capsular types associated with respiratory disorders in cattle, sheep, and goats. For the 129 isolates analyzed, both q-PCR and IHA assays exhibited nearly complete agreement for capsular types A1 (k = 0.965) and A2 (k = 0.888) and substantial agreement for A6 (k = 0.801). Despite the overall good performance of the commercial q-PCR, its effectiveness differed between the host origin of the isolates. The serotype was identified by q-PCR in 83.3 % of cattle, 77.8 % of goat, and 53.8 % of sheep isolates. Combining the results of both methods, A1 was the most prevalent in cattle and sheep (55.6 % and 22.25 %, respectively) but was not detected in goats, A2 was the most prevalent in goats (61.1 %) and the second most prevalent in cattle (16.7 %) and sheep (20.5 %). The prevalence of A6 was 7.4 %, 5.1 %, and 16.7 % in cattle, sheep, and goats, respectively. Other capsular types determined exclusively by IHA were A16 in cattle, A9 in goats, and A7, A8, A9, and A13 in sheep. Capsular type diversity was greater in sheep (H = 0.601) than in cattle (H = 0.408) and goat (H = 0.330) isolates. The commercial multiplex q-PCR is a valuable tool, alternative to IHA, for identifying isolates of capsular types A1, A2, and A6, the most frequent serotypes of M. haemolytica associated with respiratory disease in ruminants. However, when testing sheep isolates it should be complemented with immunological assays due to the wider range of serotypes implicated.


Asunto(s)
Cápsulas Bacterianas/clasificación , Enfermedades de los Bovinos/microbiología , Enfermedades de las Cabras/microbiología , Mannheimia haemolytica/clasificación , Enfermedades de las Ovejas/microbiología , Animales , Bovinos , Enfermedades de los Bovinos/epidemiología , Enfermedades de las Cabras/epidemiología , Cabras , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Ovinos , Enfermedades de las Ovejas/epidemiología , España/epidemiología
5.
mBio ; 12(3)2021 05 18.
Artículo en Inglés | MEDLINE | ID: mdl-34006665

RESUMEN

The polysaccharide capsule is a key virulence factor of Streptococcus pneumoniae There are numerous epidemiologically important pneumococcal capsular serotypes, and recent findings have demonstrated that several of them are commonly found among nonpathogenic commensal species. Here, we describe 9 nonpneumococcal strains carrying close homologs of pneumococcal capsular biosynthetic (cps) loci that were discovered during recent pneumococcal carriage studies of adults in the United States and Kenya. Two distinct Streptococcus infantis strains cross-reactive with pneumococcal serotype 4 and carrying cps4-like capsular biosynthetic (cps) loci were recovered. Opsonophagocytic killing assays employing rabbit antisera raised against S. infantis US67cps4 revealed serotype 4-specific killing of both pneumococcal and nonpneumococcal strains. An S. infantis strain and two Streptococcus oralis strains, all carrying cps9A-like loci, were cross-reactive with pneumococcal serogroup 9 strains in immunodiffusion assays. Antiserum raised against S. infantis US64cps9A specifically promoted killing of serotype 9A and 9V pneumococcal strains as well as S. oralis serotype 9A strains. Serotype-specific PCR of oropharyngeal specimens from a recent adult carriage study in the United States indicated that such nonpneumococcal strains were much more common in this population than serotype 4 and serogroup 9 pneumococci. We also describe S. oralis and S. infantis strains expressing serotypes identical or highly related to serotypes 2, 13, and 23A. This study has expanded the known overlap of pneumococcal capsular serotypes with related commensal species. The frequent occurrence of nonpneumococcal strains in the upper respiratory tract that share vaccine and nonvaccine capsular serotypes with pneumococci could affect population immunity to circulating pneumococcal strains.IMPORTANCE The distributions and frequencies of individual pneumococcal capsular serotypes among nonpneumococcal strains in the upper respiratory tract are unknown and potentially affect pneumococcal serotype distributions among the population and immunity to circulating pneumococcal strains. Repeated demonstration that these nonpneumococcal strains expressing so-called pneumococcal serotypes are readily recovered from current carriage specimens is likely to be relevant to pneumococcal epidemiology, niche biology, and even to potential strategies of employing commensal live vaccines. Here, we describe multiple distinct nonpneumococcal counterparts for each of the pneumococcal conjugate vaccine (PCV) serotypes 4 and 9V. Additional data from contemporary commensal isolates expressing serotypes 2, 13, and 23A further demonstrate the ubiquity of such strains. Increased focus upon this serological overlap between S. pneumoniae and its close relatives may eventually prove that most, or possibly all, pneumococcal serotypes have counterparts expressed by the common upper respiratory tract commensal species Streptococcus mitis, Streptococcus oralis, and Streptococcus infantis.


Asunto(s)
Cápsulas Bacterianas/clasificación , Portador Sano/microbiología , Serogrupo , Streptococcus/clasificación , Streptococcus/genética , Anciano , Anciano de 80 o más Años , Animales , Cápsulas Bacterianas/genética , Cápsulas Bacterianas/inmunología , Reacciones Cruzadas/inmunología , Humanos , Conejos , Streptococcus/inmunología , Streptococcus/aislamiento & purificación , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/inmunología , Simbiosis , Estados Unidos
6.
Vet Microbiol ; 257: 109077, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33901804

RESUMEN

Pasteurella multocida is an important cause of pneumonic pasteurellosis in small ruminants. Its prevalence was investigated in 349 pneumonic lungs from sheep (n = 197) and goats (n = 152), and genotypes of isolates were determined by capsular and lipopolysaccharide (LPS) typing as well as by virulotyping based on the detection of 12 virulence-associated genes. P. multocida was isolated from 29.4 % of sheep lungs and 13.8 % of goat lungs. A (78.5 %) and D (21.5 %) capsular types, as well as L3 (41.8 %) and L6 (57.0 %) LPS genotypes, were detected, with the A:L6 genotype being the most prevalent in both sheep (59.6 %) and goat (52.4 %) isolates. A total of 19 virulence profiles (VP) were detected, seven non-toxigenic and 12 toxigenic, which correlated with the capsular-LPS genotype. All isolates of each VP belonged to the same LPS and capsular genotype, except for one isolate of VP1. The diversity in VP was higher among toxigenic (0.29) than non-toxigenic (0.18) isolates. Moreover, the toxigenic VPs showed more diversity in their capsular-LPS genotypes, with the two main toxigenic VPs belonging to genotypes D:L3 (VP2) and A:L3 (VP3). Therefore, the abundance of toxigenic isolates among sheep and goat isolates does not seem to correspond to the expansion of a more virulent lineage associated with pneumonic pasteurellosis in small ruminants. The most prevalent genotypes among sheep isolates were the non-toxigenic VP1:A:L6 (41.4 %) and the toxigenic VP3:A:L3 (17.2 %) genotypes, whereas the most prevalent among goat isolates were the toxigenic VP2:D:L3 (33.3 %) and the non-toxigenic VP1:A:L6 (14.3 %) and VP4:A:L6 (14.3 %) genotypes. These prevalent toxigenic and non-toxigenic genotypes seem to be epidemiologically relevant in pneumonic pasteurellosis of small ruminants.


Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Genotipo , Pasteurella multocida/genética , Pasteurella multocida/patogenicidad , Pasteurelosis Neumónica/microbiología , Factores de Virulencia/genética , Animales , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Toxinas Bacterianas/biosíntesis , Variación Genética , Enfermedades de las Cabras/epidemiología , Enfermedades de las Cabras/microbiología , Cabras/microbiología , Irán/epidemiología , Lipopolisacáridos/clasificación , Lipopolisacáridos/genética , Pasteurella multocida/clasificación , Pasteurelosis Neumónica/epidemiología , Rumiantes/microbiología , Ovinos/microbiología , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/microbiología , Virulencia/genética , Factores de Virulencia/clasificación
7.
Vet Microbiol ; 255: 109021, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33667982

RESUMEN

Two serologically and molecularly non-typeable isolates of the porcine lung pathogen Actinobacillus pleuropneumoniae have been identified from diseased swine in two different continents. Genome sequencing was carried out to identify their diagnostically relevant genotypes. Both isolates are biovar 1 and encode genes for production of ApxIV and ApxII (apxIICA structural genes, and apxIBD export genes). They both possess the same novel type II capsule locus (most similar to serovar 1, but with two capsule genes not previously found in A. pleuropneumoniae) but differ in their O-Ag loci. Strain 7213384-1 from Denmark, which we propose as the reference strain for serovar 19, has a serogroup 3/6/8/15 O-Ag locus; the Canadian isolate A08-013 has a serogroup 4/7 O-Ag locus. We have expanded the second of our two previously described A. pleuropneumoniae mPCRs to include capsule gene-specific primers for definitive detection of serovars 13-14 and 16-19.


Asunto(s)
Actinobacillus pleuropneumoniae/clasificación , Cápsulas Bacterianas/clasificación , Reacción en Cadena de la Polimerasa Multiplex/métodos , Serotipificación/métodos , Actinobacillus pleuropneumoniae/genética , Cápsulas Bacterianas/química , ADN Bacteriano/genética , Genoma Bacteriano
8.
Int J Infect Dis ; 104: 580-583, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33476756

RESUMEN

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae) of serogroup 19 are mainly represented by serotypes 19A and 19F, which are associated with antimicrobial resistance and disease. The wzy gene, a component of the pneumococcal capsular locus, is the target to differentiate serotypes 19A and 19F by PCR-based capsular typing. In the last decade, allelic variants of the wzy19F gene have been described, leading to misinterpretation of capsular typing results. METHODS: A collection of 154 serotype 19F S. pneumoniae strains recovered from carriage and disease in Brazil was evaluated to identify and characterize wzy19F variant isolates. RESULTS: Eleven (7%) wzy19F variant isolates were detected and identified as belonging to ST810 (n = 10) or ST13673 (n = 1; single-locus variant of ST810). They were mostly recovered from diseased patients, susceptible to the antimicrobial agents tested (except for one multidrug-resistant strain) and did not harbor pili genes. Sequences of the wzy19F gene of these variants were identical to each other and to those previously described in Brazil, but slightly different from wzy19F variants identified in other countries. CONCLUSION: This study indicated that wzy19F variants present a geographically driven distribution and was the first to uncover phenotypic and genetic features of a wzy19F variant lineage occurring in Brazil since 1989.


Asunto(s)
Infecciones Neumocócicas/microbiología , Streptococcus pneumoniae/clasificación , Streptococcus pneumoniae/aislamiento & purificación , Alelos , Infecciones Asintomáticas , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Brasil , Proteínas Portadoras/genética , Portador Sano/microbiología , Femenino , Variación Genética , Humanos , Masculino , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa , Serogrupo , Serotipificación , Streptococcus pneumoniae/genética
9.
Vet Microbiol ; 247: 108750, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32768204

RESUMEN

Streptococcus suis is a pig pathogen and a vector of zoonotic diseases that can cause severe systemic infection in humans. S. suis can colonize the nasal cavity, tonsils, and upper respiratory, genital, and digestive tracts in healthy pigs. Here, to determine prevalence, serotype distribution, and antimicrobial susceptibility of S. suis in healthy pigs, we collected 1813 nasal cavity samples from healthy pigs raised on 17 independent farms in six Chinese provinces between 2016 and 2018. We obtained 223 S. suis isolates (12.3 %) and the antimicrobial susceptibility to a panel of 11 antimicrobial agents was measured by microbroth dilution. Most S. suis isolates (98.7 %) were resistant to at least three classes of antimicrobial agents. The optrA gene conferring resistance to oxazolidinones and phenicols was identified in the chromosome of 27 isolates and on a ∼40-kb plasmid in one isolate; to the best of our knowledge, this was the first report of plasmid-borne optrA gene in S. suis. The genetic environment of optrA showed substantial diversity and could be divided into eleven different types. Interestingly, some fragments of the 89 K pathogenicity island (PAI) were observed together with optrA in 3 isolates, which warrants further attention. Capsular serotypes of S. suis isolates were determined by multiplex PCR. Serotype 29 was the most prevalent, followed by serotype 7 and serotype 2. The presence of highly virulent serotype 2 strains may pose a threat to public health.


Asunto(s)
Antiinfecciosos/farmacología , Cápsulas Bacterianas/clasificación , Farmacorresistencia Bacteriana Múltiple/genética , Oxazolidinonas/farmacología , Infecciones Estreptocócicas/veterinaria , Streptococcus suis/efectos de los fármacos , Animales , China/epidemiología , Filogenia , Serogrupo , Infecciones Estreptocócicas/epidemiología , Streptococcus suis/genética , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/microbiología , Secuenciación Completa del Genoma , Zoonosis/epidemiología , Zoonosis/microbiología
10.
J Infect Dev Ctries ; 14(6): 606-613, 2020 06 30.
Artículo en Inglés | MEDLINE | ID: mdl-32683351

RESUMEN

INTRODUCTION: The clinical and molecular characteristics of hypervirulent Klebsiella pneumoniae (hvKp) in various provinces of China have been reported, however, there have been few reports in Hebei Province, North China. METHODOLOGY: The hvKp was identified by PCR amplification of hypervirulence-related genes, the hypermucoviscous phenotype was determined by the "string test", the drug susceptibility analysis was performed using the VITEK® 2 Compact Bacterial Identification and Monitoring System. Logistic regression was used to identify risk factors for hvKp infection. The molecular epidemiological characteristics of the strains were analyzed by pulsed-field gel electrophoresis (PFGE), and the capsular serotype of hvKp strain was detected by PCR. RESULTS: Overall, 52.21% (59/113) of K. pneumoniae isolates were hvKp, and the ratios of patients with older ages or a higher PMN cell count among hvKp infection were higher than those among classical Klebsiella pneumoniae (cKp) infection. hvKp are more susceptible to antibacterial drugs than cKp, and one ESBLs-producing hvKp strain was detected. The main capsular serotype of hvKp were K2, K57 and K1. PFGE indicated that the 59 strains of hvKp could be classified into 51 PFGE band types, forming 6 PFGE clusters. CONCLUSIONS: In this study, the detection rate of hvKp was 52.21% (59/113) identified by virulence genes. People with older ages or a higher PMN cell count are more likely to gain hvKp infection. ESBLs-producing hvKp is emerging, indicating the importance of epidemiologic surveillance and clinical awareness of this pathogen in this region.


Asunto(s)
Infecciones por Klebsiella/epidemiología , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/patogenicidad , Factores de Virulencia/genética , Anciano , Antibacterianos/farmacología , Cápsulas Bacterianas/clasificación , China/epidemiología , Farmacorresistencia Bacteriana , Monitoreo Epidemiológico , Femenino , Hospitales/estadística & datos numéricos , Humanos , Infecciones por Klebsiella/inmunología , Klebsiella pneumoniae/clasificación , Klebsiella pneumoniae/efectos de los fármacos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Serogrupo , Virulencia
11.
mBio ; 11(3)2020 05 19.
Artículo en Inglés | MEDLINE | ID: mdl-32430472

RESUMEN

Streptococcus pneumoniae (pneumococcus) is a major human pathogen producing structurally diverse capsular polysaccharides. Widespread use of highly successful pneumococcal conjugate vaccines (PCVs) targeting pneumococcal capsules has greatly reduced infections by the vaccine types but increased infections by nonvaccine serotypes. Herein, we report a new and the 100th capsule type, named serotype 10D, by determining its unique chemical structure and biosynthetic roles of all capsule synthesis locus (cps) genes. The name 10D reflects its serologic cross-reaction with serotype 10A and appearance of cross-opsonic antibodies in response to immunization with 10A polysaccharide in a 23-valent pneumococcal vaccine. Genetic analysis showed that 10D cps has three large regions syntenic to and highly homologous with cps loci from serotype 6C, serotype 39, and an oral streptococcus strain (S. mitis SK145). The 10D cps region syntenic to SK145 is about 6 kb and has a short gene fragment of wciNα at the 5' end. The presence of this nonfunctional wciNα fragment provides compelling evidence for a recent interspecies genetic transfer from oral streptococcus to pneumococcus. Since oral streptococci have a large repertoire of cps loci, widespread PCV usage could facilitate the appearance of novel serotypes through interspecies recombination.IMPORTANCE The polysaccharide capsule is essential for the pathogenicity of pneumococcus, which is responsible for millions of deaths worldwide each year. Currently available pneumococcal vaccines are designed to elicit antibodies to the capsule polysaccharides of the pneumococcal isolates commonly causing diseases, and the antibodies provide protection only against the pneumococcus expressing the vaccine-targeted capsules. Since pneumococci can produce different capsule polysaccharides and therefore reduce vaccine effectiveness, it is important to track the appearance of novel pneumococcal capsule types and how these new capsules are created. Herein, we describe a new and the 100th pneumococcal capsule type with unique chemical and serological properties. The capsule type was named 10D for its serologic similarity to 10A. Genetic studies provide strong evidence that pneumococcus created 10D capsule polysaccharide by capturing a large genetic fragment from an oral streptococcus. Such interspecies genetic exchanges could greatly increase diversity of pneumococcal capsules and complicate serotype shifts.


Asunto(s)
Cápsulas Bacterianas/química , Cápsulas Bacterianas/clasificación , Serogrupo , Streptococcus pneumoniae/clasificación , Estudios Transversales , Humanos , Sueros Inmunes , Inmunización , Fagocitosis , Vacunas Neumococicas , Polisacáridos Bacterianos/química
12.
Sci Rep ; 9(1): 19823, 2019 12 27.
Artículo en Inglés | MEDLINE | ID: mdl-31882619

RESUMEN

In children, the incidence of pneumococcal meningitis has decreased since the introduction of pneumococcal conjugate vaccine (PCV7 and PCV13). However, since the introduction of the vaccine, developed countries have seen the emergence of non-PCV13 serotypes. However, invasive pneumococcal disease (IPD) caused by PCV13-targeted serotypes still represents an important public health problem in resource-limited countries. To develop a rapid, simple, and cost-effective assay to detect serotypes of Streptococcus pneumoniae, we developed a novel loop-mediated isothermal amplification (LAMP) assay based on the sequences available for the 13 capsular types that are included in PCV13: 1, 3, 4, 5, 6 A, 6B, 7 F, 9 V, 14, 18 C, 19 A, 19 F, and 23 F. We evaluated test reactivity, specificity, sensitivity and performance, and compared the results between established LAMP and conventional PCR assays. To support its clinical use, the detection limits of the LAMP assay were evaluated using bacterial genomic DNA-spiked cerebrospinal fluid (CSF) and blood specimens. We confirmed the specificity of the LAMP assay using 41 serotypes of pneumococcal strains. The sensitivity of the LAMP assay was 10 to 100 copies per reaction, compared to 10 to 104 copies per reaction for PCR assays. The detection limits of the LAMP assay were comparable when using DNA-spiked CSF and blood specimens, as compared to using purified DNA as the template. In conclusion, a rapid and simple LAMP-based pneumococcal serotyping method has been developed. This is the first report of a LAMP method for a PCV13 serotype-specific identification assay, which could be a promising step to facilitate epidemiological studies of pneumococcal serotyping.


Asunto(s)
ADN Bacteriano/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Neumonía Neumocócica/diagnóstico , Streptococcus pneumoniae/genética , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Secuencia de Bases , Preescolar , Femenino , Humanos , Lactante , Masculino , Neumonía Neumocócica/microbiología , Sensibilidad y Especificidad , Serogrupo , Serotipificación/métodos , Streptococcus pneumoniae/clasificación , Streptococcus pneumoniae/fisiología
13.
Am J Trop Med Hyg ; 101(3): 541-548, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31392944

RESUMEN

Campylobacter jejuni is the leading bacterial cause of diarrhea worldwide. A capsular polysaccharide (CPS) conjugate vaccine is under development and requires determination of the valency. However, distribution of CPS types circulating globally is presently poorly described. We aimed to determine whether CPS type distribution in Peru differs from that in other endemic regions. We used a multiplex polymerase chain reaction (PCR) assay for the detection of CPS encoding genes capable of distinguishing all 35 CPS types on Campylobacter isolates in two prospective communities based studies conducted in cohorts of children less than 59 months of age in Peru. Results showed that CPS type HS4 complex was the most prevalent, followed by HS3 complex and HS15. Differences in CPS type for symptomatology were not statistically significant. Most subjects demonstrated repeated infections over time with different CPS types, suggesting that CPS types may confer of a level of homologous protective immunity. In this dataset, some differences in CPS type distribution were observed in comparison to other low-middle income countries. Further studies need to be conducted in endemic areas to increase our knowledge of CPS type distribution and guide vaccine development.


Asunto(s)
Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/microbiología , Campylobacter jejuni/genética , Infecciones Asintomáticas/epidemiología , Infecciones por Campylobacter/diagnóstico , Campylobacter jejuni/clasificación , Preescolar , ADN Bacteriano/genética , Diarrea/epidemiología , Diarrea/microbiología , Femenino , Humanos , Lactante , Masculino , Perú/epidemiología , Prevalencia , Estudios Prospectivos
14.
FEMS Microbiol Lett ; 366(8)2019 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-31089702

RESUMEN

Capsules are a critical virulence factor in many pathogenic Escherichia coli, of which groups 2 and 3 capsules are synthesised by the ABC transporter pathway. The well-studied forms are in group 2 and much of our knowledge of group 3 is inferred from our understanding of group 2. We analyse six group 3 gene clusters including representatives of K10, K11 and K96, and find unexpected diversity. Groups 2 and 3 both have gene clusters with terminal regions 1 and 3 containing mostly genes shared by all members of both groups, plus a central region 2, that in group 2 has the genes for synthesising the serotype-specific repeat unit. We find that in all but one case group 3 gene clusters include, in addition to serotype-specific genes, a previously unrecognised set of shared genes in region 2 that probably codes for an additional structural element. Also, the six shared genes in regions 1 and 3 of group 3 exist in two very different sequence forms. It appears that the E. coli ABC transporter capsules have a very long history, with more fundamental diversity present in group 3, but greater diversity in the exposed strongly antigenic serotype-specific component encoded by region 2.


Asunto(s)
Transportadoras de Casetes de Unión a ATP/genética , Cápsulas Bacterianas/genética , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Familia de Multigenes , Cápsulas Bacterianas/clasificación , Escherichia coli/patogenicidad , Variación Genética , Polisacáridos Bacterianos , Serogrupo , Factores de Virulencia
15.
Microbiol Spectr ; 7(2)2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30873933

RESUMEN

The surface of the Gram-positive opportunistic pathogen Streptococcus agalactiae, or group B Streptococcus (GBS), harbors several carbohydrate and protein antigens with the potential to be effective vaccines. Capsular polysaccharides of all clinically-relevant GBS serotypes coupled to immunogenic proteins of both GBS and non-GBS origin have undergone extensive testing in animals that led to advanced clinical trials in healthy adult women. In addition, GBS proteins either alone or in combination have been tested in animals; a fusion protein construct has recently advanced to human clinical studies. Given our current understanding of the antigenicity and immunogenicity of the wide array of GBS surface antigens, formulations now exist for the generation of viable vaccines against diseases caused by GBS.


Asunto(s)
Antígenos Bacterianos/inmunología , Antígenos de Superficie/inmunología , Cápsulas Bacterianas/inmunología , Infecciones Estreptocócicas/prevención & control , Vacunas Estreptocócicas/inmunología , Streptococcus agalactiae/inmunología , Animales , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Femenino , Humanos , Inmunogenicidad Vacunal , Polisacáridos Bacterianos/inmunología , Infecciones Estreptocócicas/inmunología , Streptococcus agalactiae/genética , Streptococcus agalactiae/patogenicidad , Vacunas Conjugadas , Factores de Virulencia/inmunología
16.
Arch Virol ; 164(5): 1493-1497, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30895405

RESUMEN

In the present study, we investigate the biological properties and genomic organization of virulent bacteriophage AM24, which specifically infects multidrug-resistant clinical Acinetobacter baumannii strains with a K9 capsular polysaccharide structure. The phage was identified as a member of the family Myoviridae by transmission electron microscopy. The AM24 linear double-stranded DNA genome of 97,177 bp contains 167 open reading frames. Putative functions were assigned for products of 40 predicted genes, including proteins involved in nucleotide metabolism and DNA replication, packaging of DNA into the capsid, phage assembly and structural proteins, and bacterial cell lysis. The gene encoding the tailspike, which possesses depolymerase activity towards the corresponding capsular polysaccharides, is situated in the phage genome outside of the structural module, upstream of the genes responsible for packaging of DNA into the capsid. The data on characterization of depolymerase-carrying phage AM24 contributes to our knowledge of the diversity of viruses infecting different capsular types of A. baumannii.


Asunto(s)
Acinetobacter baumannii/virología , Cápsulas Bacterianas/metabolismo , Genes Virales/genética , Myoviridae/clasificación , Myoviridae/genética , Acinetobacter baumannii/efectos de los fármacos , Cápsulas Bacterianas/clasificación , ADN Viral/genética , Farmacorresistencia Bacteriana Múltiple , Genoma Viral/genética , Microscopía Electrónica de Transmisión , Myoviridae/aislamiento & purificación , Sistemas de Lectura Abierta/genética , Análisis de Secuencia de ADN
17.
Vaccine ; 36(38): 5678-5684, 2018 09 11.
Artículo en Inglés | MEDLINE | ID: mdl-30122645

RESUMEN

BACKGROUND: Haemophilus influenzae type b (Hib) vaccine was introduced as a voluntary vaccine in December 2008 and was included in the national routine immunization program in April 2013 in Japan. Currently, no nationwide data are available to evaluate the effectiveness of Hib vaccine in Japan. METHODS: To evaluate the effectiveness of Hib vaccine in Japan, nationwide active population-based surveillance of culture-proven invasive infections caused by H. influenzae in children was performed in 2008-2017 in 10 prefectures in Japan (covering approximately 23% of the total Japanese population). Clinical data were recorded on a standardized case report form. Capsular type and antimicrobial susceptibility of the H. influenzae isolates were examined. The incidence rate ratio (IRR) and its confidence interval (CI) were calculated to compare data from 5 years before and that from after the introduction of the national routine Hib vaccine immunization program. RESULTS: During the 10-year study period, 566 invasive H. influenzae disease cases including 336 meningitis cases were identified. The average number of invasive H. influenzae disease cases among children <5 years of age during 2013-2017 decreased by 93% (IRR: 0.07, 95%CI 0.05-0.10, p < 0.001) compared with those occurring during 2008-2012. Hib strains have not been isolated from invasive H. influenzae disease cases since 2014; however, non-typeable H. influenzae and H. influenzae type f isolates have been noted as causes of invasive H. influenzae diseases among children <5 years in the post-Hib vaccine era. CONCLUSIONS: After the governmental subsidization of the Hib vaccine, invasive Hib disease cases decreased dramatically in the study population, as per our surveillance. Continuous surveillance is necessary to monitor the effectiveness of Hib vaccine and for detecting any emerging invasive capsular types.


Asunto(s)
Cápsulas Bacterianas/inmunología , Vacunas contra Haemophilus/inmunología , Haemophilus influenzae tipo b/inmunología , Programas de Inmunización , Meningitis por Haemophilus/epidemiología , Meningitis por Haemophilus/prevención & control , Vigilancia de la Población , Vacunas Conjugadas/inmunología , Antibacterianos/uso terapéutico , Cápsulas Bacterianas/clasificación , Preescolar , Femenino , Vacunas contra Haemophilus/administración & dosificación , Humanos , Lactante , Japón/epidemiología , Masculino , Meningitis por Haemophilus/inmunología , Vacunación , Vacunas Conjugadas/administración & dosificación , Resistencia betalactámica/genética , beta-Lactamasas/biosíntesis , beta-Lactamasas/genética
18.
Vet Microbiol ; 220: 83-89, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29885806

RESUMEN

Problems with serological cross-reactivity have led to development of a number of PCRs (individual and multiplex) for molecular typing of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Most of these assays were developed for detection of specific amplicons within capsule biosynthetic genes before the availability of complete sequences for the different serovars. Here we describe comparative analysis of the complete capsular loci for all 18 serovars of A. pleuropneumoniae, and development of two multiplex PCRs for comprehensive capsule typing of this important pig pathogen.


Asunto(s)
Actinobacillus pleuropneumoniae/genética , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Reacción en Cadena de la Polimerasa Multiplex/métodos , Polisacáridos Bacterianos/genética , Análisis de Secuencia , Enfermedades de los Porcinos/diagnóstico , Infecciones por Actinobacillus/diagnóstico , Infecciones por Actinobacillus/microbiología , Actinobacillus pleuropneumoniae/clasificación , Actinobacillus pleuropneumoniae/patogenicidad , Animales , Cápsulas Bacterianas/química , Serogrupo , Serotipificación , Porcinos , Enfermedades de los Porcinos/microbiología
20.
Artículo en Inglés | MEDLINE | ID: mdl-29209595

RESUMEN

Klebsiella pneumoniae is one of the most clinically relevant species in immunocompromised individuals responsible for community-acquired and nosocomial infections, including pneumonias, urinary tract infections, bacteremias, and liver abscesses. Since the mid-1980s, hypervirulent K. pneumoniae, generally associated with the hypermucoviscosity phenotype, has emerged as a clinically significant pathogen responsible for serious disseminated infections, such as pyogenic liver abscesses, osteomyelitis, and endophthalmitis, in a generally younger and healthier population. Hypervirulent K. pneumoniae infections were primarily found in East Asia and now are increasingly being reported worldwide. Although most hypervirulent K. pneumoniae isolates are antibiotic-susceptible, some isolates with combined virulence and resistance, such as the carbapenem-resistant hypervirulent K. pneumoniae isolates, are increasingly being detected. The combination of multidrug resistance and enhanced virulence has the potential to cause the next clinical crisis. To better understand the basic biology of hypervirulent K. pneumoniae, this review will provide a summarization and discussion focused on epidemiology, hypervirulence-associated factors, and antibiotic resistance mechanisms of such hypervirulent strains. Epidemiological analysis of recent clinical isolates in China warns the global dissemination of hypervirulent K. pneumoniae strains with extensive antibiotic resistance in the near future. Therefore, an immediate response to recognize the global dissemination of this hypervirulent strain with resistance determinants is an urgent priority.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Farmacorresistencia Bacteriana/fisiología , Infecciones por Klebsiella/epidemiología , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/patogenicidad , Factores de Virulencia/genética , Bacteriemia/epidemiología , Bacteriemia/microbiología , Cápsulas Bacterianas/clasificación , Cápsulas Bacterianas/genética , Proteínas Bacterianas/genética , Carbapenémicos , Infección Hospitalaria/epidemiología , Infección Hospitalaria/microbiología , Humanos , Ácidos Hidroxámicos/farmacología , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/clasificación , Absceso Hepático/epidemiología , Absceso Hepático/microbiología , Epidemiología Molecular , Filogeografía , Serogrupo , Infecciones Urinarias/epidemiología , Infecciones Urinarias/microbiología , Virulencia
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