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1.
Scand J Trauma Resusc Emerg Med ; 29(1): 46, 2021 Mar 11.
Article in English | MEDLINE | ID: mdl-33706791

ABSTRACT

BACKGROUND: Spinal stabilisation is recommended for prehospital trauma treatment. In Germany, vacuum mattresses are traditionally used for spinal stabilisation, whereas in anglo-american countries, long spine boards are preferred. While it is recommended that the on-scene time is as short as possible, even less than 10 minutes for unstable patients, spinal stabilisation is a time-consuming procedure. For this reason, the time needed for spinal stabilisation may prevent the on-scene time from being brief. The aim of this simulation study was to compare the time required for spinal stabilisation between a scoop stretcher in conjunction with a vacuum mattress and a long spine board. METHODS: Medical personnel of different professions were asked to perform spinal immobilizations with both methods. A total of 172 volunteers were immobilized under ideal conditions as well as under realistic conditions. A vacuum mattress was used for 78 spinal stabilisations, and a long spinal board was used for 94. The duration of the procedures were measured by video analysis. RESULTS: Under ideal conditions, spinal stabilisation on a vacuum mattress and a spine board required 254.4 s (95 % CI 235.6-273.2 s) and 83.4 s (95 % CI 77.5-89.3 s), respectively (p < 0.01). Under realistic conditions, the vacuum mattress and spine board required 358.3 s (95 % CI 316.0-400.6 s) and 112.6 s (95 % CI 102.6-122.6 s), respectively (p < 0.01). CONCLUSIONS: Spinal stabilisation for trauma patients is significantly more time consuming on a vacuum mattress than on a long spine board. Considering that the prehospital time of EMS should not exceed 60 minutes and the on-scene time should not exceed 30 minutes or even 10 minutes if the patient is in extremis, based on our results, spinal stabilisation on a vacuum mattress may consume more than 20 % of the recommended on-scene time. In contrast, stabilisation on a spine board requires only one third of the time required for that on a vacuum mattress. We conclude that a long spine board may be feasible for spinal stabilisation for critical trauma patients with timesensitive life threatening ABCDE-problems to ensure the shortest possible on-scene time for prehospital trauma treatment, not least if a patient has to be rescued from an open or inaccessible terrain, especially that with uneven overgrown land.


Subject(s)
Beds , Immobilization/methods , Spinal Injuries , Stretchers , Emergency Medical Services , Female , Germany , Humans , Immobilization/instrumentation , Male , Patient Simulation , Vacuum
2.
Epidemiol Infect ; 146(14): 1807-1810, 2018 10.
Article in English | MEDLINE | ID: mdl-29986784

ABSTRACT

Serosurveys have established data about the distribution of immunoglobulin G (IgG)-antibodies to pertussis toxin (PT) in various populations. We tried to detect whether small serosurveys in blood donors could serve as a simple and inexpensive means to collect information about the circulation of Bordetella pertussis. We screened every donation in 307 adult blood donors aged 19-69 years for IgG-anti-PT by standardised enzyme-linked immunosorbent assays (ELISA), and the donors were followed between 2014 and 2016 for a total of 426 person-years. When we used a vertical survey with cut-offs of 100, 62.5 and 40 IU/ml, respectively, as an indicator for recent contacts with B. pertussis, nine (2.9%), 22 (7.2%) and 54 (17.6%) of donors had IgG-anti-PT titres above the respective levels. During the horizontal observation period of 426 person years, six significant increases and two conversions were found, which lead to an estimate of 1878 contacts/100.000 person-years (1.9% per year). Median and mean IgG-anti-PT concentrations remained relatively stable from year to year during the observation period. Our findings show that small serosurveys of blood donors offer a simple and cheap method for the surveillance of B. pertussis.


Subject(s)
Blood Donors/statistics & numerical data , Epidemiological Monitoring , Immunoglobulin G/blood , Population Surveillance/methods , Whooping Cough/epidemiology , Adult , Aged , Cohort Studies , Enzyme-Linked Immunosorbent Assay , Female , Germany/epidemiology , Humans , Incidence , Male , Middle Aged , Pertussis Toxin/immunology , Seroepidemiologic Studies , Whooping Cough/microbiology , Young Adult
3.
Epidemiol Infect ; 144(4): 840-6, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26380914

ABSTRACT

Laboratory tests in adult outpatients with longer lasting coughs to identify a potential causal pathogen are rarely performed, and there is no gold standard for these diagnostic tests. While the diagnostic validity of serological tests for pertussis is well established their potential contribution for diagnosing adenovirus and influenza virus A and B infections is unclear. A sentinel study into the population-based incidence of longer lasting coughs in adults was done in Rostock (former East Germany) and Krefeld (former West Germany). A total of 971 outpatients who consulted general practitioners or internists were included. Inclusion criteria were coughing for ⩾1 week and no chronic respiratory diseases. We evaluated the performance of polymerase chain reaction (PCR) as well as IgG and IgA serology, applying a latent class model for diagnosing infections with adenovirus, B. pertussis, and influenza virus A and B. The adult outpatients first sought medical attention when they had been coughing for a median of 3 weeks. In this situation, direct detection of infectious agents by PCR had a low sensitivity. Modelling showed that additional serological tests equally improved sensitivity and specificity for diagnosis for adenovirus, B. pertussis and influenza virus A and B infections. The combination of serology and PCR may improve the overall performance of diagnostic tests for B. pertussis and also for adenovirus, and influenza virus A and B infections.


Subject(s)
Adenoviridae Infections/diagnosis , Cough/diagnosis , Diagnostic Tests, Routine/methods , Influenza, Human/diagnosis , Whooping Cough/diagnosis , Adenoviridae/isolation & purification , Adenoviridae Infections/epidemiology , Adenoviridae Infections/virology , Adolescent , Adult , Aged , Aged, 80 and over , Bordetella pertussis/isolation & purification , Cough/epidemiology , Cough/microbiology , Cough/virology , Female , Germany/epidemiology , Humans , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Influenza, Human/epidemiology , Influenza, Human/virology , Alphainfluenzavirus/isolation & purification , Betainfluenzavirus/isolation & purification , Male , Middle Aged , Models, Theoretical , Polymerase Chain Reaction , Sensitivity and Specificity , Whooping Cough/epidemiology , Whooping Cough/microbiology , Young Adult
4.
Eur J Clin Microbiol Infect Dis ; 33(5): 805-8, 2014 May.
Article in English | MEDLINE | ID: mdl-24292098

ABSTRACT

We evaluated whether the results of diagnostic polymerase chain reaction (PCR) testing combined with time since last vaccine dose could be used to monitor the effectiveness of acellular pertussis vaccines. In 258 consecutive nasopharyngeal swabs from children and adolescents with typical pertussis symptoms, 80 were positive and 178 were negative in PCR for Bordetella pertussis DNA (IS 481). Time since last vaccine dose was available for 152 patients, of which 120 were fully immunised. Among the fully vaccinated patients, the median age of 41 PCR-positive patients was 8.4 years (range 0.9-12.3) and that of 79 PCR-negative cases was 3.3 years (range 0.4-14.1) (p < 0.01). The median time since last pertussis vaccine dose was 6.05 years [95 % confidence interval (CI): 0.5-10.9] in PCR-positive cases and 2.22 years (95 % CI: 0.04-9.23) in PCR-negative cases (p < 0.001). The use of diagnostic PCR results from pertussis cases together with time since last vaccine dose permits estimates of the duration of protection after vaccination with acellular pertussis vaccines that are in keeping with more complex studies.


Subject(s)
Bordetella pertussis/isolation & purification , Pertussis Vaccine/administration & dosage , Pertussis Vaccine/immunology , Polymerase Chain Reaction/methods , Whooping Cough/prevention & control , Adolescent , Bordetella pertussis/genetics , Child , Child, Preschool , Female , Humans , Infant , Male , Time Factors , Treatment Outcome , Vaccines, Acellular/administration & dosage , Vaccines, Acellular/immunology
5.
Eur J Clin Microbiol Infect Dis ; 32(3): 421-3, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23076769

ABSTRACT

The purpose of this investigation was to test the performance of pertussis serology in diagnostic laboratories. The World Health Organization (WHO) Reference Reagent (06/142) and a sample with a low level of antibodies were sent to 200 participants of an external quality assessment (EQA) programme in Germany. The results were reported qualitatively and quantitatively, and were converted into IU/ml when possible. A total of 183 participants reported results. IgG, IgA and IgM enzyme-linked immunosorbent assays (ELISAs) with mixed antigens were used by 111, 110 and 113 participants, respectively, and 69 and 44 participants used IgG and IgA ELISAs with purified pertussis toxin (PT), respectively. IgG, IgA and IgM immunoblots were employed by 62, 63 and 11 participants, respectively. Most tests could distinguish between the positive and negative samples, but quantitative results were reported partly in non-comparable units. Only 37 % of participants used ELISAs that gave results comparable to the expected values in IU/ml and that could be interpreted according to published recommendations.


Subject(s)
Laboratory Proficiency Testing/organization & administration , Serologic Tests/standards , Whooping Cough/diagnosis , Antibodies, Bacterial/blood , Enzyme-Linked Immunosorbent Assay/methods , Germany , Humans , Immunoblotting/methods , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Quality Assurance, Health Care/organization & administration , Serologic Tests/methods
6.
Eur J Clin Microbiol Infect Dis ; 30(12): 1531-5, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21503838

ABSTRACT

Bordetella pertussis infection is mostly diagnosed by serological tests, such as by enzyme-linked immunosorbent assays (ELISAs) or by immunoblots. We compared immunoblots from five different manufacturers. Immunoblots from Euroimmun, Mikrogen, Trinity Biotech, Viramed and Virotech were used. All kits except the kit from Trinity Biotech measured IgG and IgA antibodies separately. The kits were used according to the kit inserts. Various reference preparations from the World Health Organization (WHO), the National Institute for Biological Standards and Control (NIBSC) and the Center for Biologics Evaluation and Research/Food and Drug Administration (CBER/FDA) were analysed. Patient sera with high antibody titres in ELISA, sera from patients with compatible clinical symptoms and sera from vaccinees were compared. An algorithm for interpreting quantitative values for IgG and IgA anti-pertussis toxin (PT) from in-house ELISAs was used as a reference. The sensitivity and specificity of the assays was variable when comparing the qualitative results of immunoblots with expected values of reference preparations and ELISA interpretation of patient sera. The interpretation of semi-quantitative reading of the immunoblots did not compare well to the ELISA results. Adenylate cyclase toxin as an additional antigen in two immunoblots did not effectively distinguish between infection and vaccination. Due to the lack of quantification of antibody concentrations, IgG and IgA immunoblots are of limited value in the serological diagnosis of pertussis.


Subject(s)
Antibodies, Bacterial/blood , Bordetella pertussis/immunology , Immunoblotting/methods , Immunoglobulin A/blood , Immunoglobulin G/blood , Whooping Cough/diagnosis , Antigens, Bacterial , Humans , Pertussis Vaccine/immunology , Reagent Kits, Diagnostic , Sensitivity and Specificity
7.
Eur J Clin Microbiol Infect Dis ; 30(3): 307-12, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21069406

ABSTRACT

Bordetella pertussis-specific antibodies can be detected by enzyme-linked immunosorbent assays (ELISAs) or multiplex immunoassays. Assays use purified or mixed antigens, and only pertussis toxin (PT) is specific for B. pertussis. The interpretation of results can be based on dual-sample or single-sample serology using one or two cut-offs. The EU Pertstrain group recommends that: (i) ELISAs and multiplex immunoassays should use purified non-detoxified PT as an antigen, that they should have a broad linear range and that they should express results quantitatively in International Units per millilitre (IU/ml); (ii) a single or dual diagnostic cut-off for single-serum serology using IgG-anti-PT between 50 and 120 IU/ml should be used, and diagnostic serology cannot be validly interpreted for one year after vaccination with acellular pertussis (aP) vaccines; (iii) IgA-anti-PT should only be used with indeterminate IgG-anti-PT levels or when a second sample cannot be obtained. This group discourages using: (i) other antigens in routine diagnostics, as they are not specific; (ii) micro-agglutination, due to its lack of sensitivity; (iii) immunoblots for pertussis serodiagnosis, as results cannot be quantified; (iv) other methods, such as complement fixation or indirect immunofluorescence, due to their low sensitivity and/or specificity.


Subject(s)
Antibodies, Bacterial/blood , Bordetella pertussis/immunology , Serologic Tests/methods , Whooping Cough/diagnosis , Antibodies, Bacterial/immunology , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunoassay , Pertussis Toxin/immunology , Sensitivity and Specificity , Whooping Cough/immunology
8.
J Clin Microbiol ; 48(12): 4459-63, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20943873

ABSTRACT

Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme-linked immunosorbent assays (ELISAs). We compared the performance of ELISA kits that were commercially available in Germany. Eleven measured IgG antibodies, and nine measured IgA antibodies. An in-house ELISA with purified antigens served as a reference method. Samples included two WHO reference preparations, the former Food and Drug Administration (FDA)/Center for Biologics Evaluation and Research (CBER) reference preparations, serum samples from patients with clinically suspected pertussis, and serum samples from patients having received a combined tetanus, diphtheria, and pertussis (Tdap) vaccination. Kits using pertussis toxin (PT) as an antigen showed linearity compared to the WHO Reference preparation (r2 between 0.82 and 0.99), and these kits could quantify antibodies according to the reference preparation. ELISA kits using mixed antigens showed no linear correlation to the reference preparations. Patient results were compared to results of in-house ELISAs using a dual cutoff of either ≥100 IU/ml anti-PT IgG or ≥40 IU/ml anti-PT IgG together with ≥12 IU/ml anti-PT IgA. The sensitivities of kits measuring IgG antibodies ranged between 0.84 and 1.00. The specificities of kits using PT as an antigen were between 0.81 and 0.93. The specificities of kits using mixed antigens were between 0.51 and 0.59 and were thus not acceptable. The sensitivities of kits measuring IgA antibodies ranged between 0.53 and 0.73, and the specificities were between 0.67 and 0.94, indicating that IgA antibodies may be of limited diagnostic value. Our data suggest that ELISAs should use purified PT as an antigen and be standardized to the 1st International Reference preparation.


Subject(s)
Antibodies, Bacterial/blood , Bordetella pertussis/immunology , Pertussis Vaccine/immunology , Whooping Cough/immunology , Adolescent , Adult , Enzyme-Linked Immunosorbent Assay/methods , Germany , Humans , Immunoglobulin A/blood , Immunoglobulin G/blood , Reagent Kits, Diagnostic , Sensitivity and Specificity
9.
Dtsch Med Wochenschr ; 134 Suppl 2: S86-9, 2009 Apr.
Article in German | MEDLINE | ID: mdl-19353478

ABSTRACT

Pertussis is an endemic and cyclical disease. But protection from infection and through vaccination is not long-lived. Because of high vaccination coverage among infants the age distribution of pertussis has changed: adults and adolescents are now the main reservoir for Bordetella pertussis. The current recommendations of the German Standing Vaccination Committee (STIKO) are designed to achieve life-long protection, which is feasible with acellular pertussis vaccines. Vaccination of all adults with Tdap instead of dT vaccine is recommended in many countries, because this strategy may reduce the burden of disease in adults and infants, and is cost-effective.


Subject(s)
Pertussis Vaccine/therapeutic use , Whooping Cough/immunology , Adolescent , Adult , Child , Child, Preschool , Cost of Illness , Cost-Benefit Analysis , Female , Germany , Humans , Immunization , Infant , Pregnancy , Time Factors , Whooping Cough/economics , Whooping Cough/prevention & control
10.
Eur J Clin Microbiol Infect Dis ; 28(3): 275-9, 2009 Mar.
Article in English | MEDLINE | ID: mdl-18797942

ABSTRACT

Antibody decay after a single dose of acellular pertussis vaccine containing 25 microg of pertussis toxin (PT), 25 microg of filamentous hemagglutin (FHA), and traces of pertactin (PRN) was monitored in health-care workers. Blood was sampled 4 weeks (n = 246), 1 year (n = 187), 2 years (n = 53), 3 years (n = 134), and 4 years (n = 37) after vaccination. IgG anti-PT, IgG anti-FHA and IgG anti-PRN were measured by ELISA. Peak median antibodies to PT, FHA, and PRN were 314, 785, and 84 EU/ml respectively. IgG anti-PT decreased to a median of 29% (76 EU/ml), 18% (64 EU/ml), 19% (58 EU/ml), and 20% (63 EU/ml) of the peak value after 1, 2, 3, and 4 years respectively. IgG anti-FHA decreased more slowly, but showed similar decay patterns. In German health-care workers antibodies to pertussis antigens decayed rapidly within the first year after vaccination, but remained stable after 2, 3, and 4 years. This observation may have implications for the timing of booster vaccinations in adults.


Subject(s)
Antibodies, Bacterial/blood , Health Personnel , Pertussis Vaccine/immunology , Adolescent , Adult , Enzyme-Linked Immunosorbent Assay , Germany , Humans , Immunoglobulin G/blood , Longitudinal Studies , Middle Aged , Vaccines, Acellular/immunology , Young Adult
11.
Eur J Clin Microbiol Infect Dis ; 27(2): 145-8, 2008 Feb.
Article in English | MEDLINE | ID: mdl-17965893

ABSTRACT

Bordetella pertussis is increasingly detected by real-time PCR, but most kits for extracting Bordetella-DNA from respiratory samples are not validated for this material. Respiratory clinical materials were spiked with Bordetella pertussis cells. Four ion-exchange chromatography methods from one manufacturer were used for DNA preparation. Two real-time PCRs detecting the IS481 of Bordetella pertussis and based either on a hybridisation probes format (LightCycler) or on a TaqMan format were used. All kits effectively prepared DNA for Bordetella pertussis real-time PCR. The procedures were linear over a broad range, and the lower level of sensitivity was similar. Sensitivities measured as CT values were different. Inter-assay CVs were between 6.8% and 17.3%. Two kits did not effectively remove inhibitory substances from the respiratory samples. Commercial kits are useful for preparing Bordetella pertussis DNA from respiratory samples, but even kits from one manufacturer show significant differences in effectiveness and removal of inhibitory substances.


Subject(s)
Bacteriological Techniques/methods , Bordetella pertussis/genetics , Bordetella pertussis/isolation & purification , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Polymerase Chain Reaction , Whooping Cough/diagnosis , Chromatography, Ion Exchange , DNA Transposable Elements , Humans , Reproducibility of Results , Sensitivity and Specificity
12.
Dtsch Med Wochenschr ; 131(50): 2829-34, 2006 Dec 15.
Article in German | MEDLINE | ID: mdl-17160764

ABSTRACT

BACKGROUND AND OBJECTIVE: A prospective sentinel study into the population-based incidence of pertussis in adults was done between 2001 and 2004 in Rostock (former East Germany) and Krefeld (former West Germany). PATIENTS AND METHODS: 971 outpatients, who consulted general practitioners or internists, were included. Clinical inclusion criteria were coughing for one week or more and no chronic respiratory diseases. Bordetella infection was diagnosed by PCR on nasopharyngeal swabs and ELISA for serology (IgG-anti-PT, IgA-anti-PT, IgG-anti-FHA, IgA-anti-FHA). RESULTS: We found a total of 97 cases of pertussis in this cohort. The main symptom was coughing for a median of 7-8 weeks. Population-based incidence was estimated in Krefeld at 169 cases/100000 population per year, and in Rostock at 160/100000 per year. Resource use was 120 EUR of direct medical cost and 434 euro of indirect medical cost, not including hospitalization in this study. CONCLUSIONS: Pertussis is a frequent cause of longer lasting cough in German adults, and it causes significant morbidity and costs.


Subject(s)
Whooping Cough/epidemiology , Adult , Aged , Aged, 80 and over , Comorbidity , Costs and Cost Analysis/statistics & numerical data , Cross-Sectional Studies , Family Practice/economics , Family Practice/statistics & numerical data , Female , Germany , Health Care Costs/statistics & numerical data , Humans , Incidence , Male , Middle Aged , National Health Programs/economics , National Health Programs/statistics & numerical data , Polymerase Chain Reaction , Referral and Consultation/economics , Referral and Consultation/statistics & numerical data , Sentinel Surveillance , Smoking/epidemiology , Whooping Cough/diagnosis , Whooping Cough/economics
14.
J Clin Microbiol ; 43(6): 2837-43, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15956406

ABSTRACT

Despite the widespread use of pertussis vaccines during the last decades, pertussis has remained an endemic disease with frequent epidemic outbreaks. Currently two types of vaccines are used: whole-cell vaccines (WCVs) and recently developed acellular vaccines (ACVs). The long-term aim of our studies is to assess the effect of different vaccination policies on the population structure of Bordetella pertussis and ultimately on the disease burden in Europe. In the present study, a total of 102 B. pertussis isolates from the period 1998 to 2001 from five European countries (Finland, Sweden, Germany, The Netherlands, and France) were characterized. The isolates were analyzed by typing based on variable number of tandem repeats (VNTR); by sequencing of polymorphic genes encoding the surface proteins pertussis toxin S1 and S3 subunits (ptxA and ptxC), pertactin (prn), and tracheal colonization factor (tcfA); and by fimbrial serotyping. The results reveal a relationship between geographic location and VNTR types, the frequency of the ptxC alleles, and serotypes. We have not observed a relationship between the strain characteristics we studied and vaccination programs. Our results provide a baseline which can be used to reveal changes in the B. pertussis population in Europe in the coming years.


Subject(s)
Bordetella pertussis/isolation & purification , Health Policy , Immunization Programs , Pertussis Vaccine/administration & dosage , Whooping Cough/epidemiology , Adolescent , Adult , Bacterial Proteins/genetics , Bordetella pertussis/classification , Bordetella pertussis/genetics , Child , Child, Preschool , Europe , Fimbriae Proteins , Humans , Infant , Infant, Newborn , Minisatellite Repeats/genetics , Polymorphism, Genetic , Serotyping , Vaccination , Virulence Factors/genetics , Whooping Cough/microbiology , Whooping Cough/prevention & control
15.
Infection ; 33(1): 13-7, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15750754

ABSTRACT

BACKGROUND: Acute exacerbations of chronic bronchitis (AECB) are associated with a variety of viral and bacterial infectious agents, some of which are potentially preventable by immunization. Bordetella pertussis, which causes whooping cough, has not been studied in this context. We aimed to assess the role of Bordetella infections in patients with AECB. PATIENTS AND METHODS: Patients with AECB, who presented to participating private practices in Basel, Switzerland, between October 2000 and June 2002, were evaluated by a standardized questionnaire, nasopharyngeal swabs for culture (Bordetella spp.), and PCR (Bordetella spp. and selected other respiratory pathogens) and paired blood samples for serologic diagnosis of Bordetella infection. RESULTS: A total of 26 patients (34-86 years of age) were recruited. All culture and PCR samples were negative. Serology revealed Bordetella infection in eight (31%) patients. Duration of cough was shorter in patients with Bordetella infection compared to those without Bordetella infection (mean 15 days vs 41 days, p = 0.04). Cough > or = 21 days duration was present in three (43%) of seven patients with evidence of Bordetella infection compared to 17 (94%) of 18 controls (p = 0.012). Progression to convalescence from initial to follow-up visit after 4-6 weeks was comparable between both groups. CONCLUSION: Bordetella infections appear to play a significant role in AECB and preventive measurements such as immunization with acellular pertussis vaccines should be considered. Extended investigations are necessary to confirm our preliminary and provocative findings.


Subject(s)
Bordetella Infections/epidemiology , Bordetella parapertussis , Bronchitis, Chronic/microbiology , Whooping Cough/epidemiology , Adult , Aged , Aged, 80 and over , Bordetella Infections/diagnosis , Bronchitis, Chronic/complications , Female , Humans , Male , Middle Aged , Whooping Cough/diagnosis
16.
J Clin Microbiol ; 43(1): 30-5, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15634947

ABSTRACT

Although the PCR for the detection of Bordetella pertussis is routinely performed in diagnostic laboratories, no quality assessment program has so far been described. We report on the results obtained with two external quality assessment proficiency panels sent to European laboratories. The first proficiency panel contained a series of dilutions of three previously characterized B. pertussis clinical isolates and two negative controls. No false-positive results were reported by six laboratories providing seven data sets. The reported limits of detection of the three B. pertussis strains varied between 4 and 4,000, 9 and 9,000, and 3 and 30,000 CFU/ml, respectively. The second proficiency panel, composed of a series of dilutions of reference strains of B. pertussis, B. holmesii, B. hinzii, and B. bronchiseptica, as well as negative controls, was sent to nine laboratories. One laboratory reported a negative result for a sample and reported a B. parapertussis-positive sample to be positive for B. pertussis. By using the B. pertussis-specific target gene pertactin, one laboratory detected B. pertussis with 100% specificity. All other laboratories, which used IS481-based assays, reported positive results for the samples containing B. holmesii and B. bronchiseptica, species that have occasionally been recovered from human respiratory samples. These data show that the choice of the target gene is particularly critical for the species specificity of B. pertussis PCR assays.


Subject(s)
Bordetella pertussis/isolation & purification , DNA, Bacterial/analysis , Whooping Cough/microbiology , Bordetella pertussis/genetics , Europe , False Positive Reactions , Humans , Laboratories , Polymerase Chain Reaction , Quality Control , Reproducibility of Results , Sensitivity and Specificity
17.
Lancet Infect Dis ; 2(12): 744-50, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12467690

ABSTRACT

Bordetella pertussis continues to circulate even in populations where a high vaccine coverage of infants and children is achieved. Cases in adolescents and adults are reported with increasing frequency in many countries. Adults are a reservoir for infections in very young infants, in whom pertussis may be severe and life-threatening. The salient clinical feature of pertussis in adolescents and adults is prolonged coughing, and recognising that pertussis does occur in these age groups is the most important step in its diagnosis. A laboratory diagnosis can be made by bordetella-PCR from nasopharyngeal swabs or secretions and by detection of antibodies, mainly to pertussis toxin; laboratory diagnosis is, however, not well standardised. Vaccination of adolescents and adults is now possible with acellular pertussis vaccines, which are well tolerated, immunogenic, and effective. Adolescent boosters and the vaccination of health-care workers are already included in vaccination calendars in some countries. Vaccine-recommending bodies and national health-care organisations must have locally relevant information on the transmission of pertussis from adults to infants to be able to make decisions on the advisability, feasibility, and priority for booster immunisation against pertussis.


Subject(s)
Whooping Cough , Adult , Bordetella pertussis/immunology , Controlled Clinical Trials as Topic , Humans , Infant , Pertussis Vaccine/immunology , Risk Factors , Vaccines, Acellular/immunology , Whooping Cough/diagnosis , Whooping Cough/epidemiology , Whooping Cough/immunology , Whooping Cough/prevention & control
19.
Eur J Clin Microbiol Infect Dis ; 20(12): 850-3, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11837635

ABSTRACT

To estimate the rate of asymptomatic exposure to Bordetella pertussis antigens in the German adult population and to evaluate the stability of antibodies to these antigens, antibody levels against Bordetella antigens and their variability over time were measured in German adult blood donors. One hundred forty-six regular blood donors (41 females, 105 males) were tested repeatedly for antibodies of isotypes IgG and IgA to pertussis toxin, filamentous hemagglutinin (FHA) and pertactin over a period of 2-5 years. Overall, 86% and 56% had IgG or IgA antibodies to pertussis toxin, respectively, 100% and 92% had IgG or IgA antibodies to FHA, respectively, and 83% and 93% had IgG or IgA antibodies to pertactin, respectively. One significant titer increase of both IgG anti-FHA and IgG anti-pertactin, one of IgG anti-FHA, and two of IgA anti-FHA as well as one significant decrease of IgG anti-pertussis toxin were observed during an observation period of 480.5 person-years. Antibody concentrations in men and women were not different. The data show that the level of antibodies to pertussis toxin, FHA, and pertactin remains stable over several years. Furthermore, depending on the definition of serological evidence, the rate of significant increases or decreases suggesting unrecognized exposure to Bordetella antigens was estimated to be between <0.2 and 1.0 per 100 person-years in the population studied.


Subject(s)
Antibodies, Bacterial/blood , Bordetella pertussis/immunology , Adhesins, Bacterial/immunology , Adult , Bacterial Outer Membrane Proteins/immunology , Blood Donors , Female , Hemagglutinins/immunology , Humans , Immunoglobulin A/blood , Immunoglobulin G/blood , Male , Pertussis Toxin , Virulence Factors, Bordetella/immunology
20.
Clin Diagn Lab Immunol ; 7(3): 422-6, 2000 May.
Article in English | MEDLINE | ID: mdl-10799456

ABSTRACT

Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme-linked immunosorbent assays (ELISAs). We compared the performance of five commercially available ELISA kits with the help of 65 serum specimens which were repetitively tested for evaluation of the kits. The specimens contained 20 paired serum samples from patients with clinical pertussis, 15 samples were from children vaccinated with a diphtheria-tetanus-acellular pertussis vaccine, seven specimens were taken from an interlaboratory comparison of ELISAs, and there were three reference preparations from the Food and Drug Administration's (FDA's) Laboratory of Pertussis and from our laboratory. Reference values were obtained from the FDA or from results obtained with an in-house ELISA. Commercial ELISAs were compared with respect to their reproducibility and variability, their ability to detect significant titer rises in paired serum samples, their ability to detect an immune response after vaccination, and the comparability of semiquantitative and quantitative results. Reproducibility was generally good (>89%), intra-assay variation ranged from 2.4 to 28.7%, and indeterminate results were recorded in up to 18.5% of all specimens. Most kits correctly identified the antibody response to an acellular pertussis vaccine. None of the commercial kits identified all cases of pertussis correctly, and the sensitivity ranged between 60 and 95%. All five commercial ELISAs showed great discrepancies when comparing semiquantitative results and contained obviously different antigen preparations. Our data suggest that the five commercial ELISAs tested here need further improvement and standardization.


Subject(s)
Antibodies, Bacterial/analysis , Bordetella pertussis/isolation & purification , Enzyme-Linked Immunosorbent Assay/methods , Reagent Kits, Diagnostic , Whooping Cough/diagnosis , Adolescent , Adult , Bacterial Vaccines , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay/standards , Humans , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Infant , Laboratories/standards , Middle Aged , Reproducibility of Results
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