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Applied Microbiology and Biotechnology Sep 2022Micropipette tips are currently among the most used disposable devices in bioresearch and development laboratories. Their main application is the fractionation of...
Micropipette tips are currently among the most used disposable devices in bioresearch and development laboratories. Their main application is the fractionation of solutions. New functionalities have recently been added to this device, widening their applications. This paper analyzed disposable micropipette tips as reagent holders of PCR reagents. PCR has become a prevalent and often indispensable technique in biological laboratories for various applications, such as the detection of coronavirus and other infectious diseases. A functional micropipette tip was implemented to simplify PCR analysis and reduce the contamination chances of deoxynucleotides and specific primers. This disposable device is prepared by tip coating processes of reagents, using polyvinyl alcohol polymer and additives. The coated layer is optimized to load and release PCR reagents efficiently. As a proof of concept, we show that the detection of Bordetella pertussis, the etiological agent of whooping cough whose diagnostic relies on PCR, can be quickly done using practical-functional tips. This device is an excellent example of testing the functionality and contribution of molecular diagnostic PCR tips. KEY POINTS: • Functional micropipette tips are prepared by coating with dNTPs and primers. • Functional tips are used to replace dNTPs and primers in the PCR master mix. • PCR diagnostic of Bordetella pertussis is performed using functional tips.
Topics: Bordetella pertussis; DNA Primers; DNA, Bacterial; Humans; Polymerase Chain Reaction; Whooping Cough
PubMed: 35915170
DOI: 10.1007/s00253-022-12069-9 -
Microbial Genomics Jul 2021spp. are emerging pathogens in patients with cystic fibrosis (CF) and spp. caused infections are associated with more severe disease outcomes and high intrinsic...
spp. are emerging pathogens in patients with cystic fibrosis (CF) and spp. caused infections are associated with more severe disease outcomes and high intrinsic antibiotic resistance. While conventional CF pathogens are studied extensively, little is known about the genetic determinants leading to antibiotic resistance and the genetic adaptation in spp. infections. Here, we analysed 101 spp. genomes from 51 patients with CF isolated during the course of up to 20 years of infection to identify within-host adaptation, mutational signatures and genetic variation associated with increased antibiotic resistance. We found that the same regulatory and inorganic ion transport genes were frequently mutated in persisting clone types within and between species, indicating convergent genetic adaptation. Genome-wide association study of six antibiotic resistance phenotypes revealed the enrichment of associated genes involved in inorganic ion transport, transcription gene enrichment in β-lactams, and energy production and translation gene enrichment in the trimethoprim/sulfonamide group. Overall, we provide insights into the pathogenomics of spp. infections in patients with CF airways. Since emerging pathogens are increasingly recognized as an important healthcare issue, our findings on evolution of antibiotic resistance and genetic adaptation can facilitate better understanding of disease progression and how mutational changes have implications for patients with CF.
Topics: Achromobacter; Adaptation, Physiological; Cystic Fibrosis; Denmark; Disease Progression; Drug Resistance, Multiple, Bacterial; Energy Metabolism; Genome, Bacterial; Genome-Wide Association Study; Gram-Negative Bacterial Infections; Host-Pathogen Interactions; Humans; Respiratory Tract Infections
PubMed: 34232117
DOI: 10.1099/mgen.0.000582 -
Emerging Infectious Diseases Mar 2021Pertussis is a vaccine-preventable disease, and its recent resurgence might be attributable to the emergence of strains that differ genetically from the vaccine strain....
Pertussis is a vaccine-preventable disease, and its recent resurgence might be attributable to the emergence of strains that differ genetically from the vaccine strain. We describe a novel pertussis isolate-based surveillance system and a core genome multilocus sequence typing scheme to assess Bordetella pertussis genetic variability and investigate the increased incidence of pertussis in Austria. During 2018-2020, we obtained 123 B. pertussis isolates and typed them with the new scheme (2,983 targets and preliminary cluster threshold of <6 alleles). B. pertussis isolates in Austria differed genetically from the vaccine strain, both in their core genomes and in their vaccine antigen genes; 31.7% of the isolates were pertactin-deficient. We detected 8 clusters, 1 of them with pertactin-deficient isolates and possibly part of a local outbreak. National expansion of the isolate-based surveillance system is needed to implement pertussis-control strategies.
Topics: Alleles; Austria; Bacterial Outer Membrane Proteins; Bordetella pertussis; Humans; Pertussis Vaccine; Virulence Factors, Bordetella; Whooping Cough
PubMed: 33622477
DOI: 10.3201/eid2703.202314 -
International Journal of Infectious... Oct 2019Pertussis is a highly infectious respiratory disease caused by Bordetella pertussis. Infants and young children are particularly at risk of severe and life-threatening... (Review)
Review
Pertussis is a highly infectious respiratory disease caused by Bordetella pertussis. Infants and young children are particularly at risk of severe and life-threatening disease. Infectious older individuals may transmit Bordetella pertussis to unprotected infants. Pertussis control measures have even failed in some countries with high pertussis vaccination coverage rates, leading to increased incidence rates. In 2014, this caused the World Health Organization to declare pertussis resurgent in some countries and led to recommendations regarding pertussis surveillance and national immunization programs. Despite the resurgence of pertussis, epidemiology of the disease in Southeast Asia has received little attention. In this narrative review, we describe pertussis surveillance systems, control measures, epidemiologic trends, and region-specific pertussis research in Southeast Asia. We also make recommendations for the intensification of pertussis surveillance and research in the region.
Topics: Asia, Southeastern; Bordetella pertussis; Humans; Immunization Programs; Pertussis Vaccine; Whooping Cough
PubMed: 31369823
DOI: 10.1016/j.ijid.2019.07.016 -
Clinical Infectious Diseases : An... Dec 2020To commemorate the 100th anniversary of the Nobel prize being awarded to Jules Bordet, the discoverer of Bordetella pertussis, the 12th International Bordetella...
To commemorate the 100th anniversary of the Nobel prize being awarded to Jules Bordet, the discoverer of Bordetella pertussis, the 12th International Bordetella Symposium was held from 9 to 12 April 2019 at the Université Libre de Bruxelles, where Jules Bordet studied and was Professor of Microbiology. The symposium attracted more than 300 Bordetella experts from 34 countries. They discussed the latest epidemiologic data and clinical aspects of pertussis, Bordetella biology and pathogenesis, immunology and vaccine development, and genomics and evolution. Advanced technological and methodological tools provided novel insights into the genomic diversity of Bordetella and a better understanding of pertussis disease and vaccine performance. New molecular approaches revealed previously unrecognized complexity of virulence gene regulation. Innovative insights into the immune responses to infection by Bordetella resulted in the development of new vaccine candidates. Such discoveries will aid in the design of more effective approaches to control pertussis and other Bordetella-related diseases.
Topics: Bordetella pertussis; Genomics; Humans; Pertussis Vaccine; Virulence; Whooping Cough
PubMed: 32463883
DOI: 10.1093/cid/ciaa651 -
PLoS Pathogens Feb 2023Traditionally, whooping cough or pertussis caused by the obligate human pathogen Bordetella pertussis (Bp) is described as an acute disease with severe symptoms....
Traditionally, whooping cough or pertussis caused by the obligate human pathogen Bordetella pertussis (Bp) is described as an acute disease with severe symptoms. However, many individuals who contract pertussis are either asymptomatic or show very mild symptoms and yet can serve as carriers and sources of bacterial transmission. Biofilms are an important survival mechanism for bacteria in human infections and disease. However, bacterial determinants that drive biofilm formation in humans are ill-defined. In the current study, we show that Bp infection of well-differentiated primary human bronchial epithelial cells leads to formation of bacterial aggregates, clusters, and highly structured biofilms which are colocalized with cilia. These findings mimic observations from pathological analyses of tissues from pertussis patients. Distinct arrangements (mono-, bi-, and tri-partite) of the polysaccharide Bps, extracellular DNA, and bacterial cells were visualized, suggesting complex heterogeneity in bacteria-matrix interactions. Analyses of mutant biofilms revealed positive roles in matrix production, cell cluster formation, and biofilm maturity for three critical Bp virulence factors: Bps, filamentous hemagglutinin, and adenylate cyclase toxin. Adherence assays identified Bps as a new Bp adhesin for primary human airway cells. Taken together, our results demonstrate the multi-factorial nature of the biofilm extracellular matrix and biofilm development process under conditions mimicking the human respiratory tract and highlight the importance of model systems resembling the natural host environment to investigate pathogenesis and potential therapeutic strategies.
Topics: Humans; Bordetella pertussis; Whooping Cough; Biofilms; Epithelium; Respiratory System
PubMed: 36821596
DOI: 10.1371/journal.ppat.1011193 -
BMC Pediatrics Jul 2019The diagnosis of pertussis in clinical practice continues to be a challenge worldwide as the symptoms are variable. We aimed to determine the prevalence of pertussis in...
BACKGROUND
The diagnosis of pertussis in clinical practice continues to be a challenge worldwide as the symptoms are variable. We aimed to determine the prevalence of pertussis in Chinese children irrespective of cough duration and explore the clinical characteristics of children with pertussis with different cough durations.
METHODS
This was a prospective study of children 1 month to 11 years of age with different cough durations in one large Chinese hospital. Bilateral deep posterior nasopharyngeal swabs and venepuncture for full blood count, CRP and serology and sputum were obtained when possible for investigation. E-test strips were used for testing the susceptibility of the B.pertussis isolates against erythromycin, azithromycin, sulphamethoxazole/trimethoprim, levofloxacin, amoxicillin and doxycycline. Demographic, clinical and laboratory information on culture and antimicrobial susceptibility testing was collected from children, and analyzed using SAS v.10 (SAS Institute Inc., USA).
RESULTS
After exclusions we analyzed 312 children. Ninety-seven (31.1%) children had laboratory evidence of pertussis. When grouped by cough duration, few characteristics were significant between children with and without pertussis. Of the 36 isolates, 72.2% (26/36)could not be inhibited by erythromycin and azithromycin at all. The MIC50 and MIC90 to amoxicillin were 0.75 mg/L and 1 mg/L respectively, sensitive to amoxicillin by the EUCAST points.
CONCLUSIONS
The "one-size-fits-all" clinical pertussis case definition is no longer optimal to recognize this disease. A large comprehensive study of children with all types of cough is required to make substantial inroads into increasing both the sensitivity and specificity in pertussis diagnosis, which will have a beneficial impact on public health. Amoxicillin maybe an alternative for children with marolide-resistant B.pertussis infection; however, local sensitivities are required to inform clinical practice.
Topics: Antibodies, Bacterial; Blood Cell Count; Bordetella pertussis; C-Reactive Protein; Child; Child, Preschool; China; Cough; DNA, Bacterial; Drug Resistance, Microbial; Female; Humans; Infant; Infant, Newborn; Male; Nasopharynx; Prevalence; Prospective Studies; Real-Time Polymerase Chain Reaction; Sputum; Time Factors; Whooping Cough
PubMed: 31299934
DOI: 10.1186/s12887-019-1615-3 -
Frontiers in Immunology 2019Well-adapted pathogens have evolved to survive the many challenges of a robust immune response. Defending against all host antimicrobials simultaneously would be... (Review)
Review
Well-adapted pathogens have evolved to survive the many challenges of a robust immune response. Defending against all host antimicrobials simultaneously would be exceedingly difficult, if not impossible, so many co-evolved organisms utilize immunomodulatory tools to subvert, distract, and/or evade the host immune response. spp. present many examples of the diversity of immunomodulators and an exceptional experimental system in which to study them. Recent advances in this experimental system suggest strategies for interventions that tweak immunity to disrupt bacterial immunomodulation, engaging more effective host immunity to better prevent and treat infections. Here we review advances in the understanding of respiratory pathogens, with special focus on spp., and prospects for the use of immune-stimulatory interventions in the prevention and treatment of infection.
Topics: Bordetella; Bordetella Infections; Humans
PubMed: 31921136
DOI: 10.3389/fimmu.2019.02869 -
Frontiers in Immunology 2023With the introduction of pertussis immunization for pregnant women in many countries, there has been renewed interest in the impact of whole-cell pertussis vaccine (wP)...
With the introduction of pertussis immunization for pregnant women in many countries, there has been renewed interest in the impact of whole-cell pertussis vaccine (wP) versus acellular vaccine (aP) on disease control, particularly regarding the best approach for priming. To gather evidence on this topic, we analyzed the impact of aP or wP priming on aP vaccination during pregnancy (aPpreg) in mice. Two-mother vaccination schemes were employed (wP-wP-aPpreg and aP-aP-aPpreg), and the immune response in the mothers and their offspring, as well as the protection of the offspring against challenge, were assessed. Pertussis toxin (PTx)-specific IgG responses were detected in mothers after both the second and third doses, with higher titers after the third dose, regardless of the vaccination schedule. However, a significant reduction in PTx-IgG levels was observed after 22 weeks post aPpreg immunization in mothers with the aP-aP-aPpreg scheme but not in the wP-wP-aPpreg immunized mothers. The aP-aP-aPpreg schedule triggered a murine antibody response mainly to a Th2-profile, while wP-wP-aPpreg induced a Th1/Th2 mixed profile. Both immunization schemes administered to the mothers protected the offspring against pertussis, but the wP-wP-aPpreg vaccination conferred offspring protection in all pregnancies at least up to 20 weeks after receiving the aPpreg-dose. In contrast, the immunity induced by aP-aP-aPpreg began to decline in births that occurred 18 weeks after receiving the aPpreg dose. For the aP-aP-aPpreg scheme, pups born from gestations furthest from aPpreg (+22 weeks) had lower PTx-specific IgG levels than those born closer to the application of the dose during pregnancy. In contrast, for pups born to wP-wP-aPpreg vaccinated mothers, the PTx-specific IgG levels were maintained over time, even for those born at the longest time studied (+22 weeks). It is noteworthy that only the pups born from mothers with aP-aP-aPpreg and receiving a neonatal dose of either aP or wP were more susceptible to infection than mice with only maternal immunity, suggesting interference with the induced immunity (p<0.05). However, it should be noted that mice with maternal immunity, whether vaccinated or not with neonatal doses, are better protected against colonization with than mice without maternal immunity but vaccinated with aP or wP.
Topics: Female; Humans; Pregnancy; Animals; Mice; Whooping Cough; Bordetella pertussis; Immunization; Mothers; Pertussis Toxin; Pertussis Vaccine; Immunity; Immunoglobulin G
PubMed: 37435078
DOI: 10.3389/fimmu.2023.1192119 -
The Canadian Veterinary Journal = La... Jul 2021is a promiscuous bacterium that infects a variety of species but has not been reported in free-ranging polar bears . Sera from 385 polar bears from the western Hudson...
is a promiscuous bacterium that infects a variety of species but has not been reported in free-ranging polar bears . Sera from 385 polar bears from the western Hudson Bay region, 1986 to 2017, were tested for reactivity to with enzyme-linked immunosorbent assays using anti-canine IgG and protein G as secondary reagents. Sera from bears had variable reactivity to antigens, and there was no difference among bears that had a history of coming near the town of Churchill, Manitoba, and bears that did not. Although the sources of exposure were not determined, equivalent results in both groups suggest that potential exposure to humans (aside from handling during sampling) and their animals (dogs) was not an important co-factor in sero-positivity to .
Topics: Animals; Antibodies, Bacterial; Bordetella bronchiseptica; Canada; Dogs; Manitoba; Ursidae
PubMed: 34219781
DOI: No ID Found