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Iranian Journal of Microbiology Jun 2014Rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among...
BACKGROUND AND OBJECTIVE
Rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among non-vaccinated infants for whom pertussis is life-threatening.
MATERIALS AND METHODS
Targets IS481, IS1001, BP0026 and human GAPDH gene were used to develop a multiplex real-time PCR assay based on the TaqMan technology for detection and identification of Bordetella pertussis and Bordetella parapertussis in clinical samples. A total of 121 human clinical specimens obtained within 2012-2013 were used to evaluate the multiplex real-time PCR assay. Clinical specimens were also tested for culture and conventional PCR. Sensitivity and specificity for culture, conventional PCR, and multiplex real-time PCR were measured in comparison with a clinical standard for B. pertussis infection.
RESULTS
The lower limit of detection (LLOD) of the multiplex assay was similar to the LLOD of each target in an individual assay format, which was approximately 1 genomic equivalent per reaction for IS481, IS1001 and 10 genomic equivalents per reaction for BP0026 target. When the B. pertussis assays were compared with a clinical standard for B. pertussis infection, sensitivity was 5, 59 and 89% the specificity was 100, 100 and 100% for culture, conventional PCR, and multiplex real-time PCR, respectively.
CONCLUSIONS
Developed multiplex real-time PCR offers a fast tool with high sensitivity and specificity for the diagnosis of B. pertussis and B. parapertussis infections which is suitable for implementation in a routine laboratory diagnostics.
PubMed: 25870746
DOI: No ID Found -
International Journal of Cardiology.... Jun 2021Antibiotic envelopes are being developed for cardiac implantable electronic device (CIED) wrapping to reduce the risk of infections.
INTRODUCTION
Antibiotic envelopes are being developed for cardiac implantable electronic device (CIED) wrapping to reduce the risk of infections.
METHODS
Fifteen CIED infection-associated bacterial isolates of and were used to assess biofilm formation on Hylomate® compared to titanium, silicone and polyurethane coupons pre-treated with vancomycin (400 µg/ml), bacitracin (1000 U/ml) or a combination of rifampin (80 µg/ml) plus minocycline (50 µg/ml). Scanning electron microscopy (SEM) was performed to visualize bacteria on Hylomate®.
RESULTS
There was significantly less (p < 0.05) and on Hylomate® pre-treated with vancomycin, bacitracin or rifampin plus minocycline after 24 h of incubation (≤1.00 log CFU/cm) compared with titanium, silicone or polyurethane pre-treated with vancomycin, bacitracin or rifampin plus minocycline. . biofilms were not detected (≤1.00 log CFU/cm) on pre-treated Hylomate® coupons.
CONCLUSIONS
This study showed that Hylomate® coupons pre-treated with antibiotics reduced staphylococcal and biofilm formation
PubMed: 34159252
DOI: 10.1016/j.ijcha.2021.100801 -
Journal of Clinical Microbiology Mar 1998PCR, using primers Plp1 and Plp2, was evaluated for the detection of DNA from Bordetella pertussis in bacterial strains and in nasopharyngeal samples from patients with... (Comparative Study)
Comparative Study
PCR, using primers Plp1 and Plp2, was evaluated for the detection of DNA from Bordetella pertussis in bacterial strains and in nasopharyngeal samples from patients with a cough lasting at least 7 days. The assay could detect DNA from 6 CFU of B. pertussis/10 microl of sample. Results of the PCR assay were compared with those of cultures, a determination of serum antibodies against pertussis toxin and filamentous hemagglutinin, and a clinical evaluation of 2,442 coughing episodes. The overall sensitivity of PCR was 65% (623 of 956), which was higher than the sensitivity of cultures (58%) (P < 0.001). Factors influencing the sensitivity of PCR were the interval between the onset of symptoms and sampling and the vaccination status of the patient. The specificity of PCR was 98% (1,451 of 1,486). The positive and negative predictive values were 95 and 81%, respectively. Parapertussis PCR, using primers BPPA and BPPZ, was positive in 11 of 18 culture-positive cases and was confirmed by serology in another 4 cases. In conclusion, PCR is a valuable complement to cultures and can probably replace cultures for diagnosis of B. pertussis and Bordetella parapertussis infections.
Topics: Antibodies, Bacterial; Bordetella; Bordetella Infections; Bordetella pertussis; DNA Primers; DNA, Bacterial; Diphtheria-Tetanus-Pertussis Vaccine; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Humans; Infant; Nasopharynx; Polymerase Chain Reaction; Predictive Value of Tests; Sensitivity and Specificity; Toxoids; Whooping Cough
PubMed: 9508295
DOI: 10.1128/JCM.36.3.679-683.1998 -
Innate Immunity Aug 2014Endotoxin is recognized as one of the virulence factors of the Bordetella avium bird pathogen, and characterization of its structure and corresponding genomic features...
Endotoxin is recognized as one of the virulence factors of the Bordetella avium bird pathogen, and characterization of its structure and corresponding genomic features are important for an understanding of its role in pathogenicity and for an improved general knowledge of Bordetella spp virulence factors. The structure of the biologically active part of B. avium LPS, lipid A, is described and compared to those of another bird pathogen, opportunistic in humans, Bordetella hinzii, and to that of Bordetella trematum, a human pathogen. Sequence analyses showed that the three strains have homologues of acyl-chain modifying enzymes PagL, PagP and LpxO, of the 1-phosphatase LpxE, in addition to LgmA, LgmB and LgmC, which are required for the glucosamine modification. MALDI mass spectrometry identified a high amount of glucosamine substituting the phosphate groups of B. avium lipid A; this modification was absent from B. hinzii and B. trematum. The acylation patterns of the three lipid As were similar, but they differed from those of Bordetella pertussis and Bordetella parapertussis. They were also found to be close to the lipid A structure of Bordetella bronchiseptica, a mammalian pathogen, only differing from the latter by the degree of hydroxylation of the branched fatty acid.
Topics: Amino Acid Sequence; Bordetella; Bordetella avium; Endotoxins; Fatty Acids; Genome, Bacterial; Glucosamine; Humans; Hydrolysis; Lipid A; Lipopolysaccharides; Molecular Sequence Data; Phosphates
PubMed: 24127384
DOI: 10.1177/1753425913506950 -
Journal of Clinical Microbiology Feb 2019Molecular methods offer superior sensitivity and specificity and reduce testing turnaround time from days to hours for detection of and In this study, we evaluated the...
Molecular methods offer superior sensitivity and specificity and reduce testing turnaround time from days to hours for detection of and In this study, we evaluated the performance of the automated PCR-based Aries Assay, which detects both and directly from nasopharyngeal swab specimens. The limits of detection (LoDs) were 1,800 CFU·ml for and 213 CFU·ml for The assay detected 16/18 unique / strains. Of 71 potentially cross-reacting organisms, 5 generated false positives in 1/6 replicates; none of 6 additional spp. were erroneously detected. Specimens were stable at 20 to 25°C for at least 10 h, at 4 to 8°C for 10 days, and at temperatures not exceeding -70°C for 6 months. Of 1,052 nasopharyngeal specimens from patients with suspected pertussis, 3.0% ( = 32) were positive and 0.2% ( = 2) were positive. Combining these data with Aries Assay data from 57 nasopharyngeal samples with previously confirmed or data and with data from 50 contrived samples, the proportions of positive and negative agreement of the respective Aries assays with the reference assays were 97.1% and 99.0% for and 100% and 99.7% for The Aries Assay provides accurate detection and distinction of and infections within 2 h. (This study has been registered at ClinicalTrials.gov under registration no. NCT02862262.).
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Automation, Laboratory; Bordetella Infections; Bordetella parapertussis; Bordetella pertussis; Child; Child, Preschool; Female; Humans; Infant; Infant, Newborn; Male; Middle Aged; Molecular Diagnostic Techniques; Nasopharynx; Polymerase Chain Reaction; Prospective Studies; Sensitivity and Specificity; Time Factors; Young Adult
PubMed: 30518543
DOI: 10.1128/JCM.01471-18 -
Jundishapur Journal of Microbiology Nov 2014Pertussis is a respiratory and contagious disease which is mostly caused by Bordetella pertussis and B. parapertussis. It usually spreads from person to personduring the...
BACKGROUND
Pertussis is a respiratory and contagious disease which is mostly caused by Bordetella pertussis and B. parapertussis. It usually spreads from person to personduring the incubation or catarrhal phase of the disease. Despite of large-scale vaccination, whooping cough is still an endemic disease with several outbreaks.
OBJECTIVES
The aim of this study was to determine the prevalence of pertussis and identify its causative agents, B. pertussis or B. parapertussis, from specimens collected from Iranian patients from 2004 to 2008.
PATIENTS AND METHODS
Nasopharyngeal swab samples from 347 suspected pertussis cases were collected from 18 provinces of Iran. The patients were in different age groups and were either unvaccinated or vaccinated for pertussis with whole cell vaccine (WCV). Bacterial culture, agglutination tests and quantitative PCR (qPCR) targeting IS481 and IS1001 for B. pertussis and B. parapertussis were done for every specimen, respectively.
RESULTS
The results showed that seven nasopharyngeal swab samples (2%) were positive for B. pertussis (1.7%) and B. parapertussis (0.3%) by culture and agglutination test and 30 patients had positive qPCR test results (9%).
CONCLUSIONS
Despite the fact that bacterial culture is the golden standard for the detection of B. pertussis, direct detection of bacteria from nasopharyngeal specimens can be performed by a rapid qPCR assay. In this study, high percentage of positive qPCR cases may indicate that the patients might have recovered from pertussis following antibiotic treatment before samples were collected. Rapid detection by qPCR could be important for immediate diagnosis and treatment of patients with pertussis.
PubMed: 25774274
DOI: 10.5812/jjm.12421 -
MBio Aug 2018, , and share highly homologous virulence factors and commonly cause respiratory infections in mammals; however, their host specificities and disease severities differ,...
, , and share highly homologous virulence factors and commonly cause respiratory infections in mammals; however, their host specificities and disease severities differ, and the reasons for this remain largely unknown. Adenylate cyclase toxin (CyaA) is a homologous virulence factor that is thought to play crucial roles in infections. We herein demonstrate that CyaAs function as virulence factors differently between / and CyaA bound to target cells, and its enzyme domain was translocated into the cytosol similarly to CyaA. The hemolytic activity of CyaA on sheep erythrocytes was also preserved. However, in nucleated target cells, CyaA was phosphorylated at Ser, which constitutes a motif (RSXpSXP [pS is phosphoserine]) recognized by the host factor 14-3-3, resulting in the abrogation of adenylate cyclase activity. Consequently, the cytotoxic effects of CyaA based on its enzyme activity were markedly attenuated. CyaA carries the 14-3-3 motif, indicating that its intracellular enzyme activity is abrogated similarly to CyaA; however, CyaA has Phe instead of Ser, and thus, was not affected by 14-3-3. In addition, CyaA impaired the barrier function of epithelial cells, whereas CyaA did not. Rat infection experiments suggested that functional differences in CyaA are related to differences in pathogenicity between / and , , and are bacterial respiratory pathogens that are genetically close to each other and produce many homologous virulence factors; however, their host specificities and disease severities differ, and the reasons for this remain unknown. Previous studies attempted to explain these differences by the distinct virulence factors produced by each species. In contrast, we indicated functional differences in adenylate cyclase toxin, a homologous virulence factor of The toxins of and presumably were inactivated by the host factor 14-3-3 after phosphorylation in target cells, whereas the toxin was not inactivated because of the lack of the phosphorylation site. This is the first study to show that 14-3-3 inactivates the virulence factors of pathogens. The present results suggest that pathogenic differences in are attributed to the different activities of adenylate cyclase toxins.
Topics: 14-3-3 Proteins; Adenylate Cyclase Toxin; Animals; Bordetella Infections; Bordetella bronchiseptica; Bordetella parapertussis; Bordetella pertussis; Disease Models, Animal; Epithelial Cells; Erythrocytes; Hemolysis; Phosphorylation; Protein Binding; Protein Processing, Post-Translational; Protein Transport; Rats; Sheep; Virulence Factors
PubMed: 30154257
DOI: 10.1128/mBio.00628-18 -
Journal of Clinical Microbiology Dec 2020Detection of and using molecular methods is sensitive and specific with a short turnaround time compared to other diagnostic methods. In this multicenter study, we...
Detection of and using molecular methods is sensitive and specific with a short turnaround time compared to other diagnostic methods. In this multicenter study, we compared the performance of the Simplexa Bordetella Direct kit to those of other molecular assays in detecting and differentiating and in nasopharyngeal swab specimens. The limits of detection (LODs) were 150 CFU/ml or 3 fg/μl of DNA for and 1,500 CFU/ml or 10 fg/μl of DNA for A total of 1,103 fresh and residual frozen specimens from eight clinical sites were tested. Combining the data from individual clinical sites using different comparative assays, the overall positive percent agreement (PPA) and negative percent agreement (NPA) for were 98.7% and 97.3%, respectively. The overall PPA and NPA for were 96.7% and 100%, respectively. For prospective fresh specimens, the overall PPA and NPA for both targets were 97.7% and 99.3%, respectively. For retrospective frozen specimens, the overall PPA and NPA for both targets were 92.6% and 93.2%, respectively. The percentage of invalid results was 1.0%. A cross-reactivity study using 74 non- bacterial species and five yeast species revealed that the Simplexa Bordetella Direct kit was 100% specific. The hands-on time and assay run time of the Simplexa Bordetella Direct kit are favorable compared to those of other commercial and laboratory-developed tests. In summary, the Simplexa Bordetella Direct kit has a performance comparable to those of other molecular assays for the detection of and .
Topics: Bordetella; Bordetella Infections; Bordetella parapertussis; Bordetella pertussis; Humans; Nasopharynx; Prospective Studies; Retrospective Studies; Whooping Cough
PubMed: 33055187
DOI: 10.1128/JCM.01041-20 -
Molecular Microbiology Feb 2017Nicotinamide adenine dinucleotide (NAD) is produced via de novo biosynthesis pathways and by salvage or recycling routes. The classical Bordetella bacterial species are...
Nicotinamide adenine dinucleotide (NAD) is produced via de novo biosynthesis pathways and by salvage or recycling routes. The classical Bordetella bacterial species are known to be auxotrophic for nicotinamide or nicotinic acid. This study confirmed that Bordetella bronchiseptica, Bordetella pertussis and Bordetella parapertussis have the recycling/salvage pathway genes pncA and pncB, for use of nicotinamide or nicotinic acid, respectively, for NAD synthesis. Although these Bordetellae lack the nadA and nadB genes needed for de novo NAD biosynthesis, remarkably, they have one de novo pathway gene, nadC, encoding quinolinate phosphoribosyltransferase. Genomic analyses of taxonomically related Bordetella and Achromobacter species also indicated the presence of an 'orphan' nadC and the absence of nadA and nadB. When supplied as the sole NAD precursor, quinolinate promoted B. bronchiseptica growth, and the ability to use it required nadC. Co-expression of Bordetella nadC with the nadB and nadA genes of Paraburkholderia phytofirmans allowed B. bronchiseptica to grow in the absence of supplied pyridines, indicative of de novo NAD synthesis and functional confirmation of Bordetella NadC activity. Expression of nadC in B. bronchiseptica was influenced by nicotinic acid and by a NadQ family transcriptional repressor, indicating that these organisms prioritize their use of pyridines for NAD biosynthesis.
Topics: Bacterial Proteins; Biosynthetic Pathways; Bordetella; Genes, Bacterial; Mutation; NAD; Pentosyltransferases; Quinolinic Acid
PubMed: 27783449
DOI: 10.1111/mmi.13566 -
PloS One 2022Pertussis or whooping cough is a vaccine-preventable, highly contagious, respiratory illness caused by Bordetella pertussis or Bordetella parapertussis. Infants and...
BACKGROUND
Pertussis or whooping cough is a vaccine-preventable, highly contagious, respiratory illness caused by Bordetella pertussis or Bordetella parapertussis. Infants and young children have remained most susceptible to pertussis-related morbidity and mortality. The aim of this study was to investigate pertussis infection and analyze the associated factors involved in the occurrence of the cases.
METHODS
Community-based case-control was conducted in Dahena district, Northwest Ethiopia, from March 27-April 30, 2019. All cases ages 1-18 years old were identified by using the clinical standard case definition of pertussis adopted from World Health Organization (WHO). Data was collected using a structured questionnaire via face-to-face interviews. The data collected was cleaned, coded and entered into Epi info version 7.2.1.0 and exported to SPSS version 23 for statistical analysis. Bivariable and multivariable logistic regression analysis were employed to identify predictors. Factors with a p-value of < 0.05 were considered as independent risk factors of pertussis infection in multivariable logistic regression analysis.
RESULT
A total of 122 pertussis cases were enrolled from the Azila cluster of the Dahena district. Of these figures, 64 (52.5%) were females. The overall attack rate (AR) of pertussis cases in the cluster was 8.6/10000 population. The sex-specific AR of females was 8.9/10000 population. The multivariable logistic regression analysis showed that; being unvaccinated 4.17 (AOR, 4.17, 95% CI, 1.914-9.091), contact to cases 2.93 (AOR: 2.93, 95% CI 1.223-6.996), and living in a house with no window 2.6 (AOR: 2.6(95% CI 1.071 to 6.322) were the independent significantly risk factors for pertussis infection.
CONCLUSION
The contributing factor for pertussis infection was associated with case-contact, living in the house without windows and being unvaccinated. Wag Hemra Zone and Dahena district health office should encourage the vaccination activities of the cluster health center and awareness for the community should be practiced to limit disease transmission.
Topics: Adolescent; Case-Control Studies; Child; Child, Preschool; Disease Outbreaks; Ethiopia; Female; Humans; Infant; Logistic Models; Male; Risk Factors; Vaccination; Whooping Cough
PubMed: 35143575
DOI: 10.1371/journal.pone.0263708