Role of PCR in the diagnosis of pertussis infection in infants: 5 years' experience of provision of a same-day real-time PCR service in England and Wales from 2002 to 2007
| Autor: | Nita Doshi, Timothy G. Harrison, Robert George, Natasha S. Crowcroft, John Duncan, Norman K. Fry, David J. Litt, Karen S. Wagner, Oceanis Tzivra, Elizabeth Miller |
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| Rok vydání: | 2009 |
| Předmět: |
Adult
Microbiology (medical) Bordetella pertussis Pediatrics medicine.medical_specialty Time Factors Adolescent Bordetella Whooping Cough Pertussis toxin Polymerase Chain Reaction Sensitivity and Specificity Microbiology Bordetella parapertussis law.invention Young Adult law medicine Humans Child Polymerase chain reaction Whooping cough Aged Bordetella holmesii Wales biology business.industry Infant General Medicine Middle Aged biology.organism_classification medicine.disease Real-time polymerase chain reaction England Child Preschool business |
| Zdroj: | Journal of Medical Microbiology. 58:1023-1029 |
| ISSN: | 1473-5644 0022-2615 |
| Popis: | As part of an enhanced surveillance programme for pertussis in England and Wales, a real-time PCR service for the detection ofBordetella pertussiswas introduced for infants aged ≤6 months admitted to a paediatric intensive care unit or paediatric ward with a respiratory illness compatible with pertussis. Two real-time fluorescent resonance energy transfer hybridization probe LightCycler (Roche Diagnostics) PCR assays were used. One (designed in-house) targeted the pertussis toxin S1 promoter (ptxA-pr), and included an internal process control to test for sample inhibition and reagent performance. The other (already published) targeted the insertion element IS481. The analytical sensitivities of the assays were 100 and 10 fg per reaction for theptxA-pr and IS481PCRs, respectively. TheptxA-pr assay was specific forB. pertussis, whilst the IS481PCR also showed some cross-reactivity withBordetella holmesiiand the type strain ofBordetella parapertussis. From April 2002 to March 2007, 848 samples were received from 774 patients and DNA was extracted. Of 824 samples that were suitable for testing, 183 (22.2 %) had evidence ofBordetellainfection (18.9 %ptxA-pr and IS481; 3.3 % IS481only), 621 (75.4 %) were negative and 20 (2.4 %) were inhibitory for the PCR. Within the targeted age group of ≤6 months, most patients (130/138) with evidence ofBordetellaspp. by PCR were ≤3 months old. The overall percentage increase in laboratory-confirmed cases due to PCR compared with culture for the 5 year period described ranged from 9 to 26 % per year (mean 19 %). Real-time PCR is an invaluable tool both for enhanced epidemiological surveillance and for the provision of a rapid diagnosis of pertussis where results can affect patient and contact management. |
| Databáze: | OpenAIRE |
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