Seroepidemiology of whooping cough in the Czech Republic: estimates of incidence of infection in adults.

OBJECTIVES: Despite widespread vaccination programmes, pertussis remains a significant health burden in many countries. Low awareness of the disease, the high rate of asymptomatic cases in adults and difficulties with diagnosis could explain the under-reporting of pertussis. The lack of data on actual incidence constitutes an obstacle for public health authorities for the implementation of a vaccination strategy against pertussis in adults. The aim of this study was to evaluate the seroprevalence of antibodies against Bordetella pertussis infection in adults and to estimate the actual incidence of the disease compared with the reported incidence. STUDY DESIGN: Prospective, multicentre seroprevalence study. METHODS: The study was conducted in 2000 adult subjects aged ≥18 years who had not received pertussis vaccination within the last 5 years. All enrolled subjects provided a blood sample for serum testing of IgG antibodies against pertussis toxin, performed by enzyme-linked immunosorbent assay, to indicate if they had an acute infection or if they had been infected with pertussis within the last 12 months or earlier. Results were validated in accordance with the guidelines of the European Sero-epidemiology Network 2 and were expressed in ESEN units/ml. RESULTS: A positive concentration of anti-pertussis toxin antibodies, indicating previous pertussis infection, was demonstrated in 39.9% (n = 799) of all study subjects, and 0.40% (n = 8) of subjects had a concentration suggestive of a recent infection (within the last 12 months). The highest antibody seroprevalence was observed in subjects aged 18-29 years (1.46%). No cases of acute infection were detected. CONCLUSIONS: During the study period, the reported incidence of pertussis in the adult population was 0.84/100,000 inhabitants. Based on the seroprevalence results from this study, it is estimated that the actual incidence of pertussis could be as much as 699/100,000 inhabitants. The actual incidence of pertussis in adults in the Czech Republic could therefore be at least 200-fold higher than the reported incidence. These findings support the need for pertussis vaccination in the adult population.

[Multiple-locus variable number tandem repeat analysis of Bordetella pertussis strains collected in the Czech Republic in 1967-2015: spread of a variant adapted to the population with a high vaccination coverage]. / Multiple-locus variable number tandem repeat analýza souboru kmenu Bordetella pertussis v CR z období 1967-2015: rozsírení varianty adaptované na proockovanou populaci.

AIM: To perform multiple-locus variable number tandem repeat analysis (MLVA) of B. pertussis strains from the collection of the National Reference Laboratory for Diphtheria and Pertussis (NRL/DIPE), National Institute of Public Health (NIPH), Prague. The study strains were isolated from clinical specimens collected mostly in the Czech Republic over a nearly 50-year period from 1967 to 2015 (June). The isolates from three periods characterized by different vaccination strategies and trends in pertussis are compared for genetic diversity and distribution of MLVA types (MT). Based on the results obtained, the suitability for use of MLVA in the analysis of epidemic outbreaks of B. pertussis in the Czech Republic is considered. MATERIAL AND METHODS: DNA samples extracted from B. pertussis strains included in the present study were examined by MLVA using the standard protocol. Data were processed by means of the eBURST algorithm and the calculation of the Simpson diversity index (DI) was used for the statistical analysis. Data were analyzed as a whole and also separately for strains from the three periods: 1967-1980, 1990-2007, and 2008-2015 (June). RESULTS: Fourteen different MT were detected in the study strains, with three of them not being reported before. The most common MTs were MT27 and MT29. MT29 was predominant in 1967-1980 while MT27 was the most prevalent in 1990-2007 and 2008-2015 (June). The DI was the lowest (0.49) in 2008-2015 (June), and comparably higher DIs were calculated for the two previous periods (i.e. 0.667 for 1967-1980 and 0.654 for 1990-2007). CONCLUSION: MLVA revealed a decrease in genetic diversity and shifts in MT distribution of B. pertussis strains isolated from clinical specimens in the Czech Republic from 1967 to 2015 (June). These shifts in the Czech Republic can be characterized as a progressive increase in global MTs at the expense of the locally unique ones. The most common MT, similarly to other geographical areas with long-term high vaccination coverage, is MT27. The results of MLVA of 136 B. pertussis strains can provide a background for using this method in molecular epidemiological analysis of smaller groups of strains.

[Antigenic variability of Bordetella pertussis strains isolated in 1967-2010 in the Czech Republic--possible explanation for the rise in cases of pertussis?]. / Antigenní variabilita kmenu Bordetella pertussis izolovaných v letech 1967-2010 v Ceské republice--mozné vysvetlení vzestupu nemocnosti pertusí?

OBJECTIVE: Comparison of antigenic structures of Bordetella pertussis (B. pertussis) strains isolated from 1967 to 2010 in the Czech Republic. MATERIAL AND METHODS: Seventy strains of B. pertussis were referred to the National Reference Laboratory (NRL) for Pertussis and Diphtheria within the surveillance of pertussis from all over the Czech Republic (CR) between 1967 and 2010. To study the strains, the analysis was performed of the genome sequences encoding the surface immunogenic structures--the pertussis toxin S1 subunit gene (ptxA), pertactin gene region 1 (prnA), type 3 fimbriae gene (fim3)--and pertussis toxin promoter (ptxP) responsible for the regulation of the production of pertussis toxin. RESULTS: For the study set of B. pertussis strains, the sequencing analysis revealed changes in all genomic regions studied. The isolates from three periods differ in the allelic profile. In period I (19671978) with the use of whole cell pertussis vaccine (wP), the following two profiles were the most common: ptxP(1), ptxA(2), prnA(1), fim3(1) and ptxP(1), ptxA(1), prnA(3), fim3(1). In period 2 (19902007) with the switch to acellular pertussis vaccine (aP), the most common profile was: ptxP(3), ptxA(1), prnA(2), fim3(2). Period 3 (20082010) with the use of aP was characterized by the predominance of the following two profiles which had never been found in period 1: ptxP(3), ptxA(1), prnA(2), fim3(2) and ptxP(3) ptxA(1), prnA(2), fim3(1). CONCLUSIONS: Sequencing of the genomic regions ptxP, ptxA, prnA, and fim3 of B. pertussis strains isolated in the CR between 1967 and 2010 confirmed changes in the allelic variants of these regions. The incidence of strains carrying the new allelic variants was increasing after 1995 at the expense of those carrying the original variants. The study results can be interpreted as a partial genetic escape of pathogenic strains of B. pertussis beyond the reach of the pertussis vaccines.

[Real-time PCR detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae DNA in clinical specimens]. / Detekce DNA Neisseria meningitidis, Haemophilus influenzae a Streptococcus pneumoniae v klinickém materiálu metodou real-time PCR.

The study aim was to implement a molecular real-time polymerase chain reaction (PCR) assay recommended by the CDC (Centers for Disease Control and Prevention) for the detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical (culture negative) specimens from patients with suspected invasive bacterial disease. Clinical specimens are referred to the National Reference Laboratory (NRL) for Meningococcal Infections, Unit for Airborne Bacterial Infections, Centre for Epidemiology and Microbiology, National Institute of Public Health from various regions of the Czech Republic. Clinical specimens are, in particular, cerebrospinal fluid, anti-coagulated blood or serum and, exceptionally, post-mortem specimens. The NRL has implemented molecular diagnosis of these bacterial pathogens involved in meningitis and sepsis from clinical specimens since 1999. The first diagnostic method was semi-nested PCR followed by electrophoretic analysis. In 2014, a molecular qualitative real-time PCR assay was implemented.