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

CHRU de Tours, Service de Bactériologie-Virologie, Hôpital Bretonneau, Tours, France.
Journal of clinical microbiology (Impact Factor: 3.99). 09/2011; 49(11):3943-6. DOI: 10.1128/JCM.00335-11
Source: PubMed


We evaluated the performances of 4 commercial real-time PCR kits for Bordetella pertussis IS481 sequence detection in nasopharyngeal aspirates by comparison with an in-house real-time PCR assay. Among them, the Simplexa
Bordetella pertussis/parapertussis assay (Focus Diagnostics), the SmartCycler Bordetella pertussis/parapertussis assay (Cepheid), and Bordetella R-gene (Argene) present sensitivities over 90%. One kit proved unsuitable for routine clinical

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Available from: Fabien Garnier, Mar 12, 2014
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    • "Applied protocols were proved to have sufficient stability and reproducibility of the results than the home made protocols. Real time PCR method has recently been reported with a high sensitivity ratio [20]. However, our PCR signal based method had high agreement with ELISA in results in our study, since all these positive cases had no vaccination history [21]. "
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    ABSTRACT: Even with high coverage of vaccination programs, Bordetella pertussis is still reported in various countries. It causes a high rate of mortality and morbidity in infants while it could be asymptomatic in adults. At the present study, we are going to evaluate the frequency of B. pertussis among received specimens. This cross-sectional study was performed on 138 children under one year who were suspected to have whooping cough from October 2008 to March in 2011. Nasopharyngeal dacron and rayon swabs and sera were used for PCR and serology respectively. The mean age of the subjects was 1.9± 0.9 months. PCR was positive in 12 cases; ELISA was in agreement with PCR results except in one case that showed the specific antibody at borderline limit. The rate of reported positive results showed that pertussis not only is still present in the community, but the number of the asymptomatic cases who are able to transmit the disease may be considerable.
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    ABSTRACT: 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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    ABSTRACT: In recent years, quantitative real-time PCR tests have been extensively developed in clinical microbiology laboratories for routine diagnosis of infectious diseases, particularly bacterial diseases. This molecular tool is well-suited for the rapid detection of bacteria directly in clinical specimens, allowing early, sensitive and specific laboratory confirmation of related diseases. It is particularly suitable for the diagnosis of infections caused by fastidious growth species, and the number of these pathogens has increased recently. This method also allows a rapid assessment of the presence of antibiotic resistance genes or gene mutations. Although this genetic approach is not always predictive of phenotypic resistances, in specific situations it may help to optimize the therapeutic management of patients. Finally, an approach combining the detection of pathogens, their mechanisms of antibiotic resistance, their virulence factors and bacterial load in clinical samples could lead to profound changes in the care of these infected patients.
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