Preparation of Bordetella pertussis DNA from respiratory samples for real-time PCR by commercial kits

Riffelmann, Marion; Schmetz, J.; Bock, Susan; Koenig, C. H. Wirsing von

doi:10.1007/s10096-007-0403-4

Cited by 7 publications

(4 citation statements)

References 8 publications

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“…During our proficiency program, the NRC included three human B. bronchiseptica isolates, all harboring the IS481 sequence and all detected by all eight laboratories (one laboratory did not provide results for QC9). These data confirm the results obtained in a previous study of the preparation of B. pertussis DNA from respiratory samples for real-time PCR (19).…”

Section: Discussionsupporting

confidence: 91%

Proficiency Program for Real-Time PCR Diagnosis of Bordetella pertussis Infections in French Hospital Laboratories and at the French National Reference Center for Whooping Cough and other Bordetelloses

et al. 2009

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With the support of a ministerial program for innovative and expensive technologies, dedicated to the economic evaluation of laboratory diagnosis of pertussis by real-time PCR, external quality assessment for real-time IS481 PCR was carried out. Coordinated by the National Centre of Reference of Pertussis and other Bordetelloses (NCR), this study aimed to harmonize and to assess the performances of eight participating microbiology hospital laboratories throughout the French territory. Between January 2006 and February 2007, 10 proficiency panels were sent by the NCR (ascending proficiency program), representing a total of 49 samples and including eight panels to analyze and evaluate the global sensitivity and specificity of real-time PCR, one to assess the limit of detection, and one to evaluate nucleic acid extraction methods. As part of the descending proficiency program, extracted DNA from clinical samples was sent by the eight participating laboratories in different panels and analyzed by the NCR. In the ascending proficiency analysis, the sensitivity and specificity of the real-time PCR methods were 92.2% and 94.3%, respectively. The limit of detection of the different methods ranged between 0.1 and 1 fg/l (0.2 to 2 CFU/l). The nucleic acid extraction methods showed similar performances. During the descending proficiency analysis, performed with 126 samples, the result of the NCR for 15 samples (11.9%) was discordant with the result obtained by the source laboratory. Despite several initial differences, harmonization was easy and performances were homogeneous. However, the risk of false-positive results remains quite high, and we strongly recommend establishment of uniform quality control procedures performed regularly.

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Section: Discussionsupporting

confidence: 91%

Proficiency Program for Real-Time PCR Diagnosis of Bordetella pertussis Infections in French Hospital Laboratories and at the French National Reference Center for Whooping Cough and other Bordetelloses

et al. 2009

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“…For manual extraction, column purification kits are recommended (62) because of their yield and ability to remove inhibitors. Ion-exchange chromatography methods perform well for extraction of DNA, although their effective removal of inhibitors varies (63).…”

Section: Pcr Assaysmentioning

confidence: 99%

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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“…There are home methods for the extraction of DNA, which are gradually being replaced by commercial methods. The latter are based on the use of ion exchange resins or magnetic separation using silica particles [46]. Neither of these methods is validated for the extraction of DNA from respiratory samples [46].…”

Section: Pcr Assaysmentioning

confidence: 99%

“…The latter are based on the use of ion exchange resins or magnetic separation using silica particles [46]. Neither of these methods is validated for the extraction of DNA from respiratory samples [46]. There are studies demonstrating that, in general, the different methods are suitable for the extraction of DNA from these samples.…”

Section: Pcr Assaysmentioning

confidence: 99%

Update on Epidemiology, Diagnosis, and Treatment of Pertussis

Hozbor¹

2018

Pertussis - Disease, Control and Challenges

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Pertussis, commonly known as whooping cough, is one of the most common vaccinepreventable infections. In adolescents and adults, infection may result in a protracted cough and is occasionally associated with substantial morbidity. In children and particularly infants, morbidity is more often substantial and the disease may be fatal. Two types of vaccines against pertussis exist: whole-cell vaccines (wP), developed in the 1940s, containing the entire inactivated Bordetella pertussis organism, and acellular vaccines (aP) constituting of 1-5 purified bacterial proteins. The aPs were developed in the 1970s in order to diminish the adverse effects that could occur in the wP vaccinations. In many industrialized countries, aP replaced the wP formulations; however, wPs are still used for primary vaccination doses in developing countries. The massive use of both types of vaccines significantly reduced the morbidity and mortality associated with the disease; nevertheless, pertussis is still an important public health problem. In fact, pertussis incidence has increased in many countries, with large sustained epidemics occurring most notably in developed countries that only use acellular vaccine for all the doses included in the calendar. This chapter focuses on some recent developments in the areas of epidemiology, diagnosis, and treatment of pertussis.

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Preparation of Bordetella pertussis DNA from respiratory samples for real-time PCR by commercial kits

Cited by 7 publications

References 8 publications

Proficiency Program for Real-Time PCR Diagnosis of Bordetella pertussis Infections in French Hospital Laboratories and at the French National Reference Center for Whooping Cough and other Bordetelloses

Proficiency Program for Real-Time PCR Diagnosis of Bordetella pertussis Infections in French Hospital Laboratories and at the French National Reference Center for Whooping Cough and other Bordetelloses

Laboratory Diagnosis of Pertussis

Update on Epidemiology, Diagnosis, and Treatment of Pertussis

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