Evaluation of Four Commercial Real-Time PCR Assays for Detection of Bordetella spp. in Nasopharyngeal Aspirates

Lanotte, Philippe; Plouzeau, Chloé; Burucoa, Christophe; Grelaud, C; Guillot, Sophie; Guiso, Nicole; Garnier, Fabien

doi:10.1128/jcm.00335-11

Cited by 33 publications

(18 citation statements)

References 13 publications

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“…Several commercial assays are available for detection of B. pertussis and B. parapertussis (135)(136)(137)(138). Most assays target IS481 and IS1001 in a real-time multiplex PCR format; examples are the assays available from Elitech, Eragen, and Focus Diagnostics (135), Cepheid (136), and Diagenode (137).…”

Section: Pcr Assaysmentioning

confidence: 99%

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Laboratory Diagnosis of Pertussis

2015

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SUMMARYThe introduction of vaccination in the 1950s significantly reduced the morbidity and mortality of pertussis. However, since the 1990s, a resurgence of pertussis has been observed in vaccinated populations, and a number of causes have been proposed for this phenomenon, including improved diagnostics, increased awareness, waning immunity, and pathogen adaptation. The resurgence of pertussis highlights the importance of standardized, sensitive, and specific laboratory diagnoses, the lack of which is responsible for the large differences in pertussis notifications between countries. Accurate laboratory diagnosis is also important for distinguishing between the several etiologic agents of pertussis-like diseases, which involve both viruses and bacteria. If pertussis is diagnosed in a timely manner, antibiotic treatment of the patient can mitigate the symptoms and prevent transmission. During an outbreak, timely diagnosis of pertussis allows prophylactic treatment of infants too young to be (fully) vaccinated, for whom pertussis is a severe, sometimes fatal disease. 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. Here we review current approaches to the diagnosis of pertussis and discuss their limitations and strengths. In particular, we emphasize that the optimal diagnostic procedure depends on the stage of the disease, the age of the patient, and the vaccination status of the patient.

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Section: Pcr Assaysmentioning

confidence: 99%

“…All assays contain an internal control, and some assays also use an extraction control (Focus, Argene). Comparisons of these assays were performed with in-house real-time PCR assays for detection of B. pertussis (135)(136)(137). The sensitivities of the commercial assays were found to range from 96% to 98% compared to the inhouse assays.…”

Section: Pcr Assaysmentioning

confidence: 99%

Laboratory Diagnosis of Pertussis

2015

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“…Susceptibility to ampicillin (10 µg), erythromycin (15 µg Detection of IS481was carried out using an inhouse reference real-time PCR method as previously described [10] until August 2012. After that time, validated commercial kits were used for the detection of both IS481 and IS1001, i.e., Simplexa Bordetella pertussis/parapertussis (FOCUS Diagnostics Cypress, California, USA) until March 2013 and Bordetella pertussis/parapertussis triplex (Bio-Evolution, Brysur-Marne, France) until September 2013, following the manufacturer's protocols [11]. B. pertussis ATCC 9797 DNA was used as positive control for the in-house realtime PCR.…”

Section: Identification Serotyping and Susceptibility Testing Of Ismentioning

confidence: 99%

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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“…This will be discussed in more detail later in this minireview. IS1001 is found in B. parapertussis at approximately 20 copies per genome (35) and has been used in multiplex PCR with IS481 to detect and differentiate B. pertussis and B. parapertussis (14,16,29,31). IS1001 is also present in isolates of B. bronchiseptica (4 of 73 human-derived isolates tested by Tatti et al [31] and 26 of 38 animal-derived isolates tested by van der Zee et al [34]) at 1 to 7 copies per genome (34 (Table 2).…”

Section: Gene Targets In Bordetella Naatsmentioning

confidence: 99%

Towards Improved Accuracy of Bordetella pertussis Nucleic Acid Amplification Tests

Loeffelholz

2012

J Clin Microbiol

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In many clinical microbiology laboratories, nucleic acid amplification tests such as PCR have become the routine methods for the diagnosis of pertussis. While PCR has greatly increased the ability of laboratories to detect Bordetella pertussis infections, it has also been associated with false-positive results that can, given the tendency of B. pertussis to cause outbreaks, result in unnecessary and costly control measures. The species specificity of Bordetella gene targets and their number of copies per genome greatly impact the performance characteristics of nucleic acid amplification tests for B. pertussis . It is crucial that laboratorians recognize these characteristics, to limit false-positive test results and prevent pseudo-outbreaks.

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Evaluation of Four Commercial Real-Time PCR Assays for Detection of Bordetella spp. in Nasopharyngeal Aspirates

Cited by 33 publications

References 13 publications

Laboratory Diagnosis of Pertussis

Laboratory Diagnosis of Pertussis

Pertussis in north-central and northwestern regions of Algeria

Towards Improved Accuracy of Bordetella pertussis Nucleic Acid Amplification Tests

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