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Semiquantitative Polymerase Chain Reaction Enzyme Immunoassay for the Diagnosis of Pertussis
Authors:R. C. Matthews  N. Golbang  W. M. Brück  D. Owen  A. Bailey  V. Weston  J. R. Kerr
Affiliation:(1) Pertussis Reference Laboratory, University Department of Medical Microbiology, Clinical Sciences Building, Central Manchester Healthcare Trust, Oxford Road, Manchester M13 9WL, UK e-mail: dorene@labmed.cmht.nwest.nhs.uk, GB;(2) Virology Unit, Clinical Sciences Building, Central Manchester Healthcare Trust, Oxford Road, Manchester M13 9WL, UK, GB;(3) Department of Microbiology and Public Health Laboratory, University Hospital, Queen's Medical Centre, Nottingham NG7 2UH, UK, GB
Abstract: The two most commonly used targets for diagnosis of pertussis by the polymerase chain reaction have been the pertussis toxin promoter and the repeated insertion sequence IS481. A comparative assessment of these primers was performed on routinely collected nasopharyngeal swabs, stored at –20  °C, using novel semiquantitative enzyme immunoassays. Both sets of primers behaved similarly with bacterial suspensions, and the 17 culture-positive nasopharyngeal swabs were also positive with the pertussis toxin promoter primers, with one exception, which had been subject to prolonged storage. Significantly more of the 69 culture-negative swabs were positive with the pertussis toxin promoter primers (n=36) than with the IS481 primers (n=18). To determine the effect of inhibitors, a comparative assessment of three primer pairs against human DNA (β-globin and glyceraldehyde-3-phosphate dehydrogenase) was also performed.
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