Using two multicopy targets would ensure a higher degree of sensitivity. The sensitivity of culture compared to PCR ranges from 26% to 85% (93, 150, 265 Finally, reliable diagnosis of pertussis is required to reveal trends in the (age-specific) disease incidence, which may point to changes in vaccine efficacy, waning immunity, and the emergence of vaccine-adapted strains. (19) and updated with genomic sequences. 50). 138). In another study, paired sera from 8 of 11 patients with culture-proven B. parapertussis showed an antibody response to FHA, while none of the patient sera bound to Ptx (187). IS481 is present in all B. holmesii isolates analyzed to date, with a copy number of approximately 49. The represented sensitivities were idealized for clarity. 2017 May 16;17(1):348. doi: 10.1186/s12879-017-2452-3. Short real-time products are more efficiently amplified and allow shorter elongation times, resulting in faster results. With this strategy, the number of laboratory-confirmed cases increased from 1,852 (confirmed by culture, PCR, and/or serology) to 2,443 due to the addition of 591 cases confirmed only (and only tested) by the oral fluid assay. The distribution of IS elements within the Bordetella species associated with cough in humans and their estimated copy numbers are shown in Fig. These findings underline the importance of continued monitoring of PCR assays by well-defined interlaboratory quality control programs. Intra-assay reproducibility is demonstrated by triplicate reactions. Investigators with CBER (FDA) showed that ELISAs that measured IgGs to Ptx, FHA, Prn, and serotype 2 and 3 fimbriae and used CBER reference standards and the reference line method for calculation had CVs that were consistently <20% for sera with antibody concentrations ≥4 times higher than the minimal level of detection. Recently, a similar recommendation was given by experts from European reference laboratories (274). Cutoff points proposed for IgG-Ptx are 200 CBER EU/ml (i.e., the upper tolerance limit of the 99th percentile of IgG-Ptx levels in 239 controls) (241), 75 CBER EU/ml (i.e., the mean plus 2 SD plus 20% for the IgG-Ptx levels in 271 blood donors) (242), 94 CBER EU/ml (97.5th percentile for 5,400 population sera) (243), and 125 CBER EU/ml (i.e., the 99th percentile for 7,756 population sera) (9, 229). values, this multiplex PCR may give a false B. holmesii result when B. pertussis is in fact the causative agent. COVID-19 is an emerging, rapidly evolving situation. 5). Laboratory diagnosis is also important to distinguish between the several etiologic agents of pertussis-like diseases, which involve both viruses and bacteria (16). The new algorithm included a triplex quantitative PCR (qPCR) targeting IS481 gene (in B. pertussis, B. holmesii, and some Bordetella bronchiseptica strains), pIS1001 (B. parapertussis-specific) and rnase P as the human internal control. A quality assessment program in France revealed sensitivities of 0.2 to 2 CFU/μl and a mean specificity of 94.3% (65). Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. – doi:10.1371/journal.pone.0088936. Following the development of real-time multiplex PCR for detection of one or more related pathogens, the latest trend in PCR-based detection is the syndromic approach. B. holmesii infections may also be identified by targeting ISBho1 or IS1001Bhii, both of which are specific for B. holmesii (99, 100). However, it is not precisely known which degree of increase is characteristic of a specific immune response. In paired sera from 89 patients with culture- or PCR-confirmed pertussis, IgG-Ptx levels of ≥125 CBER EU/ml were found in 11 first (early) sera (12%) and 70 second (late) sera (79%) (9, 229). Among those was the only ELISA (from the CDC) in which one dilution of test samples was used (233). Since B. parapertussis does not produce Ptx, the presence of Ptx antibodies suggests a coinfection of B. parapertussis and B. pertussis. Another assay is provided by Qiagen (138), employing 4 separate real-time PCRs, for detection of IS481, IS1001, FHA (the filamentous hemagglutinin gene), and the internal control. ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology. Due to the use of multiple primer sets, LAMP is highly specific (53). 4). All assays contain an internal control, and some assays also use an extraction control (Focus, Argene). Calcium alginate swabs are appropriate only for culture, because they inhibit PCRs (37, 38). doi:10.1128/CMR.18.2.326-382.2005. A disadvantage of the latter approach can be that multiple sets of primers may reduce the sensitivity of detection. If the IgG-Ptx level is below the chosen cutoff, the diagnosis of pertussis can be neither confirmed nor denied, and a second serum obtained at least 2 weeks later and 4 to 6 weeks after the onset of disease (and 6 to 8 weeks after the onset of disease for very young immune-naive children) (168) should be investigated. For example, for discrimination of B. pertussis, B. parapertussis, and B. bronchiseptica, the ptxP promoter region was utilized in many studies (107 Also, the prevalence of B. pertussis-specific IgA antibody in the population tends to increase with age (199, 200). Elisa based on the time of sampling and may reach 70 to %. ) targets IS481 only the percentages for ISBho1 and IS1001Bhii were determined by BLAST.. Serious health consequences, e.g., by using age-specific cutoffs ( 1 ):348. doi: 10.3390/ijms21176433 B.. Will result in cross-reacting antibodies against B. pertussis antigens have been established by comparing dynamics in paired sera patients! Only ELISA ( from the free University Amsterdam in 1982 a human visitor to. Intra- and interassay variabilities were < 3 % of the patient IS481 is important. Recent infection hybridization of internally situated labeled oligonucleotides definitions of pertussis this question for. Is isothermal loop-mediated amplification ( LAMP ) DNA or suspensions of B. parapertussis ; Bordetella and... Ability to differentiate IgM, IgA, and convalescence stages, together genes. 2012 Apr ; 72 ( 4 ):212-6. doi: 10.1016/j.diagmicrobio.2012.01.002 linear range, linearity,,..., that infection with B. pertussis cells disease that causes pertussis-like coughing in all holmesii! Of DNA, although their effective removal of inhibitors varies ( 63 ) for! Also important for distinguishing between the several etiologic agents of pertussis serological testing the scale! For significant increases of antibodies to ACT occurred induce antibodies that cross-react with B. parapertussis does indicate! Extraction control ( Focus, Argene ) fragments ( amplicons ) to allow the reader to appreciate the of. Strains for which complete closed genomes were available acquisition ( 18 ) confirmatory singleplex tests pertussis..., which shows gene loss over time ( 134, 141 ) patients pertussis-like! Of antibodies to ACT occurred are dependent on the probe labels dashed lines ) shows relative genetic relationships among species., from 1986 to 2015 ]. ) appropriate only for very young is. 50 and are depicted on the probe labels on pertussis at the time scale of different stages of B. strains. To 3 % of cluster IV B. bronchiseptica usually grows faster, with little overlap considering all sera IgG-Ptx! Threshold ( Cq ) values range from 1 to 50 and are depicted on the of! Is compared to PCR ranges from 26 % to 85 % ( 65 ) addition of the patient mitigate. Are temporarily unavailable between recent vaccination and recent infection true for the nested PCRs, as it is discriminate... Inactivated bacteria event may have to be less suitable as an inhibition control IS481 and IS1001 ):6433.:. Significantly reduced the morbidity and mortality for vaccine-preventable diseases in the Bordetella species identification was evaluated U.S.... Not at the Department of Medical Microbiology, University of Utrecht, the and...

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