Real-Time LightCycler PCR for Detection and Discrimination of
            <i>Bordetella pertussis</i>
            and
            <i>Bordetella parapertussis</i>

Kösters, Katrin; Reischl, Udo; Schmetz, J.; Riffelmann, Marion; König, Carl Heinz Wirsing von

doi:10.1128/jcm.40.5.1719-1722.2002

Cited by 74 publications

(49 citation statements)

References 29 publications

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“…This is in agreement with several studies reporting that B. pertussis was the main etiological agent of pertussis [35]. A negative result for B. parapertussis is consistent with the low rate of isolation of this bacterium previously reported in infants [36].…”

Section: Pertussis Confirmed Case Patientssupporting

confidence: 81%

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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Section: Pertussis Confirmed Case Patientssupporting

confidence: 81%

Pertussis in north-central and northwestern regions of Algeria

Benamrouche

Maamar

Lazri

et al. 2016

J Infect Dev Ctries

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“…Several PCR protocols have been developed in recent years for different targets in the genome of B. pertussis like an insertion sequence IS481, pertussis toxin gene promoter (ptxA-Pr), pertussis toxin S1 subunit (ptxS1), porin gene, pertactin filaments hemagglutinin gene and adenylate cyclase (14,15,20,21). Many laboratories use the insertion sequence IS481 in PCR assays to determine the presence of B. pertussis DNA.…”

Section: Discussionmentioning

confidence: 99%

“…The high sensitivity (70-90%) and specificity (86-100%), along with a shorter response time to results, low risk of contamination and ease of performance, makes RT-PCR an alternative method for the diagnosis of many diseases, among them, the pertussis (13)(14)(15).…”

Section: Introductionmentioning

confidence: 99%

Implementation and Assessment of the Use of Real-Time PCR in Routine Diagnosis for Bordetella pertussis Detection in Brazil

Leite

Blanco

Melo

et al. 2013

Arch Pediatr Infect Dis

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Background: Bordetella pertussis is the causative agent of pertussis. In Brazil, laboratory diagnosis of pertussis is based on the culture. In 2010, was standardized the Real-Time PCR TaqMan® in routine diagnosis. Objectives: The aim of this study was to evaluate the impact achieved with the introduction of RT-PCR for the routine diagnosis of pertussis and to compare with the results obtained from culture. Patients and Methods: 4,697 samples of nasopharyngeal secretions collected from suspected pertussis cases and/or contacts were analyzed for RT-PCR and culture, from January 2008 until the end of December 2011. Results: According to the results obtained from the samples 6.9% were culture/RT-PCR positive, 14.8% were positive only for RT-PCR and 0.2% only for culture. Negative samples for both techniques was 3,622 (77.1%) and 1.0% were inconclusive for RT-PCR. Conclusions:The implementation of RT-PCR in routine diagnosis resulted in an increase in laboratory confirmation by almost three times. The RT-PCR assay does not intend to replace the culture technique, but to promote an improvement in the diagnosis of pertussis.

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mentioning

confidence: 99%

“…Insertion sequences include IS481 in B. pertussis, B. holmesii, and, occasionally B. bronchiseptica, and IS1001 in B. parapertussis and, occasionally, B. bronchiseptica and B. holmesii. These sequences have been reported as target sequences in several PCR protocols (5,10,13,15).In the present study, the new artus Bordetella LC PCR Kit (QIAGEN Hamburg GmbH, Germany), which includes a multiplex PCR system consisting of four independent PCRs for qualitative detection and differentiation of B. pertussis, B. parapertussis, B. bronchiseptica, and the heterologous internal control (IC), was evaluated. After we tested these methods for analytical sensitivity and specificity, clinical specimens were investigated.…”

mentioning

confidence: 99%

Detection and Differentiation of Bordetella spp. by Real-Time PCR

Koidl

Bozic

Burmeister³

et al. 2007

J Clin Microbiol

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Molecular detection of Bordetella pertussis DNA is a sensitive and specific method for the rapid diagnosis of pertussis. In this study, a new molecular assay for the detection and differentiation of Bordetella spp. based on automated DNA extraction and real-time PCR was evaluated. The analytical sensitivity of the new assay was determined by Probit analysis of serial dilutions of both cloned PCR products IS481 and IS1001 and cell suspensions of B. pertussis, B. parapertussis, and B. bronchiseptica. The specificity was analyzed by testing a number of pathogens producing respiratory infections. Moreover, a total of 92 clinical samples were investigated. The results were compared to those obtained by an in-house assay based on manual DNA extraction, followed by real-time PCR and detection of IS481. The analytical sensitivity of the new assay for the detection of IS481 and IS1001 was determined to be 2.2 and 1.2 genome equivalents/l, respectively. The analytical sensitivity for the detection of B. pertussis, B. parapertussis, and B. bronchiseptica was determined to be 1.6, 1.0, and 2.7 genome equivalents/l, respectively. When clinical specimens were tested with the new assay, 46 of 92 were found to be positive for Bordetella DNA. With the in-house assay, 45 samples tested positive. The new molecular assay proved to be suitable for the rapid diagnosis of pertussis in the routine diagnostic laboratory.Bordetella pertussis is the agent responsible for producing pertussis, also called whooping cough. B. parapertussis, B. bronchiseptica, and B. holmesii may also cause a pertussis-like respiratory disease but with a milder clinical course (7). Pertussis occurs worldwide, with an increased incidence among young unvaccinated infants. Neonates and elderly patients may show an atypical course of disease (2, 4). A rapid and reliable diagnostic method is thus essential for appropriate treatment and prophylaxis. Culture has been considered the gold standard for detection of B. pertussis but often lacks sensitivity, and a minimum of 4 days may be required to obtain results (1).Today, PCR has been shown to be a sensitive and specific method for the rapid diagnosis of pertussis (3,6,11). In the Bordetella genome, there are repetitive insertion sequences present in a high copy number that may be useful for the amplification of different bacterial strains. Insertion sequences include IS481 in B. pertussis, B. holmesii, and, occasionally B. bronchiseptica, and IS1001 in B. parapertussis and, occasionally, B. bronchiseptica and B. holmesii. These sequences have been reported as target sequences in several PCR protocols (5,10,13,15).In the present study, the new artus Bordetella LC PCR Kit (QIAGEN Hamburg GmbH, Germany), which includes a multiplex PCR system consisting of four independent PCRs for qualitative detection and differentiation of B. pertussis, B. parapertussis, B. bronchiseptica, and the heterologous internal control (IC), was evaluated. After we tested these methods for analytical sensitivity and specificity, clinical speci...

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Real-Time LightCycler PCR for Detection and Discrimination of Bordetella pertussis and Bordetella parapertussis

Cited by 74 publications

References 29 publications

Pertussis in north-central and northwestern regions of Algeria

Pertussis in north-central and northwestern regions of Algeria

Implementation and Assessment of the Use of Real-Time PCR in Routine Diagnosis for Bordetella pertussis Detection in Brazil

Detection and Differentiation of Bordetella spp. by Real-Time PCR

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