Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis

Antila, Mia; He, Qiushui; Jong, Caroline de; Aarts, Ingrid; Verbakel, Harold; Bruisten, Sylvia M.; Keller, Suzanne M.; Haanperä, Marjo; Mäkinen, Johanna; Eerola, Erkki; Viljanen, Matti K.; Mertsola, Jussi; Zee, Anneke van der

doi:10.1099/jmm.0.46331-0

Cited by 74 publications

(36 citation statements)

References 45 publications

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“…It was underlined that real-time ptxA-Pr PCR was less sensitive but more specific than IS481 PCR (24), since IS481 PCR can detect Bordetella holmesii species and some Bordetella bronchiseptica and B. pertussis spp. This may not impact substantially on test performance, since the incidence of B. holmesii respiratory infections seems to be very low, as is that of B. bronchiseptica, which infects mostly immunosuppressed individuals (2,11). In the present study, we confirmed that ptxA-Pr real-time PCR is less sensitive than IS481 PCR and demonstrated that performing both ptxA-Pr and IS481 PCR does not improve diagnostic performance.…”

Section: Discussionsupporting

confidence: 73%

Comparison of Serological and Real-Time PCR Assays To Diagnose Bordetella pertussis Infection in 2007

et al. 2008

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Bacterial culture for diagnosing pertussis infection has high specificity but poor sensitivity and is slow. Highly sensitive real-time PCR assays and single-serum pertussis serology have been developed to overcome these limitations, but there are few data available on the relative sensitivities and specificities of such assays for pertussis diagnosis. Using data on 195 participants (>7 years old) from an epidemiological study, we assessed the sensitivity, specificity, and performance (Youden index) for pertussis diagnosis of the pertussis toxin enzyme-linked immunosorbent assay (using single and paired serology) and of real-time PCR assays (using the IS481 and ptxA-Pr targets). All available diagnostic information (clinical and laboratory) was pooled to serve as the gold standard. Single serology was the most efficient diagnostic test (Youden index, 0.57 to 0.58), with relatively high sensitivity (>64%) and high specificity (>90%), independent of the cutoff level. IS481 PCR performance was superior to that of ptxA-Pr PCR, and it was the second-most-efficient tool (Youden index, 0.30). Performing both ptxA-Pr and IS481 PCRs did not improve diagnostic performance. The greatest test efficiency (Youden index, 0.69 to 0.74) was achieved when single-serum serology was used in combination with IS481 or ptxA-Pr PCR or paired serology. Combining single serology with one PCR or paired serology increased the sensitivity with an associated limited decrease in specificity. The most specific tests for diagnosis of pertussis were single serology and ptxA-Pr PCR, and the most sensitive diagnostic tool was the combination of IS481 PCR with single serology.Currently there is no satisfactory gold standard technique for laboratory confirmation of a pertussis infection. Although culture is highly specific, sensitivity is low and declines with the duration of illness, and the method may take up to 7 days to provide a result. Using culture alone to diagnose pertussis is likely to lead to underreporting of pertussis cases (8). Serological assays, using one or two serum samples, have been developed to improve the sensitivity of the pertussis diagnosis, but they have a lower specificity than culture (3,9,20,25). PCR techniques were developed to overcome these limitations, but despite a consensus meeting in 2005 (24), these techniques still require further standardization and optimization. To date there are few reports (8) in which the sensitivities and specificities of these different techniques are compared using a population exposed to pertussis.The objective of this study was to compare the sensitivities and specificities of the current most widely used techniques to diagnose pertussis in exposed populations, namely, two PCR methods (IS481 and ptxA-Pr targets) and single and paired serology for detection of antipertussis toxin (PT) antibodies in the serum of the patients using enzyme-linked immunosorbent assay (ELISA) with two different cutoffs (two-or fourfold change for paired serology and Ն100 or Ն125 ELISA units [EU]/ml for single...

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Section: Discussionsupporting

confidence: 73%

Comparison of Serological and Real-Time PCR Assays To Diagnose Bordetella pertussis Infection in 2007

et al. 2008

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“…A few assays using more than one target per reaction have been published (1,6,10,22,23,26). The region of IS481, pIS1001, and hIS1001 targeted in our assay is sensitive, with LLODs of Յ1 Bordetella genomic equivalent per reaction ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Novel Multitarget Real-Time PCR Assay for Rapid Detection of Bordetella Species in Clinical Specimens

et al. 2011

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A novel multitarget real-time PCR (RT-PCR) assay for the rapid identification of Bordetella pertussis, B. parapertussis, and B. holmesii was developed using multicopy insertion sequences (ISs) in combination with the pertussis toxin subunit S1 (ptxS1) singleplex assay. The RT-PCR targets for the multiplex assay include IS481, commonly found in B. pertussis and B. holmesii; IS1001 of B. parapertussis; and the IS1001-like sequence of B. holmesii. Overall, 402 Bordetella species and 66 non-Bordetella species isolates were tested in the multitarget assay. Cross-reactivity was found only with 5 B. bronchiseptica isolates, which were positive with IS1001 of B. parapertussis. 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 all targets. A total of 197 human clinical specimens obtained during cough-illness outbreak investigations were used to evaluate the multitarget RT-PCR assay. The multiplex assay results from 87 clinical specimens were compared to the individual RT-PCR assay and culture results. The multitarget assay is useful as a diagnostic tool to confirm B. pertussis infections and to rapidly identify other Bordetella species. In conclusion, the use of this multitarget RT-PCR approach increases specificity, while it decreases the amount of time, reagents, and specimen necessary for RT-PCRs used for accurate diagnosis of pertussis-like illness.

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“…IS481 is present in higher copy numbers in B. pertussis (80 to 193 copies per genome) than in B. holmesii (28 to 36 copies per genome) and B. bronchiseptica (≤1 copy per genome) [20]. However, a retrospective study of 11,319 PCR assays obtained in 1992-2003 using an IS481 target in Finnish and Dutch patients with suspected pertussis did not detect B. holmesii despite finding 1,856 PCR-positive isolates [21]. The results of a similar four-year prospective study using an IS481 target and conducted from 2007 to 2011 in 599 Tunisian infants did not detect B. holmesii in a total of 126 confirmed cases [22].…”

Section: Laboratory Findingsmentioning

confidence: 94%

Pertussis in north-central and northwestern regions of Algeria

Benamrouche

Maamar

Lazri

et al. 2016

J Infect Dev Ctries

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Introduction: Pertussis outbreaks continue to occur in many countries despite high vaccination coverage. Under-diagnosed cases in adolescents and adults may result in increased transmission to infants, who are at risk of severe pertussis. Additional measures to protect both groups should be considered. Methodology: Nasopharyngeal samples and sera were collected from patients and household contacts with clinically suspected pertussis. Diagnoses were confirmed by culture, real-time polymerase chain reaction (PCR), and serology. Bordetella pertussis isolates were characterized by antimicrobial sensitivity and fimbrial serotyping. Results: Of 392 participants, 134/248 patients (54%) and 66/144 contacts (45.8%) had confirmed pertussis infections. B. parapertussis was not detected. All B. pertussis isolates were sensitive to the antibiotics tested, and all expressed the Fim3, not the Fim2, fimbrial serotype. Most patients (81.2%) were <6 months (51.8% of whom were <3 months) of age; 77.6% were unvaccinated, and most positive contacts were mothers 20-40 years of age. Conclusions: Despite high vaccination coverage, pertussis is circulating in Algeria. Most infections occur in unvaccinated infants <6 months of age, with mothers as the main source of infection. An adolescent/adult booster should be considered. Adoption of sensitive and specific laboratory tests would improve pertussis diagnosis and surveillance.

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Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis

Cited by 74 publications

References 45 publications

Comparison of Serological and Real-Time PCR Assays To Diagnose Bordetella pertussis Infection in 2007

Comparison of Serological and Real-Time PCR Assays To Diagnose Bordetella pertussis Infection in 2007

Novel Multitarget Real-Time PCR Assay for Rapid Detection of Bordetella Species in Clinical Specimens

Pertussis in north-central and northwestern regions of Algeria

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