Development of a rapid diagnostic test for pertussis: direct detection of pertussis toxin in respiratory secretions.

Monoclonal antibodies (MAb) were produced against the specific Bordetella pertussis antigen pertussis toxin (PT). In preliminary studies, one MAb (IB12) was selected and used in an enzyme-linked dot blot immunoassay to evaluate the ability of the method to detect known amounts of PT in control experiments and to test its potential for direct detection of PT in nasopharyngeal secretion (NP) specimens from patients with confirmed cases of whooping cough. The dot blot assay was able to detect PT at levels as low as 10 ng per dot in either buffer or control NP specimens. The assay demonstrated specificity, reacting only with dot blots of whole B. pertussis and not Bordetella bronchiseptica, Bordetella parapertussis, or other bacterial strains. In preliminary studies, NP aspirate, swab, and wash specimens were compared. The specimen of choice was found to be the NP aspirate, for which 100% positive results were found in the assay. These initial studies suggest that the dot blot immunoassay in which a MAb is used for direct detection of PT in NP specimens may be useful as a rapid diagnostic test for pertussis.

Activation analysis with standards containing two or more active nuclides A computerized method of calculation involving decay and gamma spectral resolution.

The resolution of the gamma spectra of activities induced in materials by fast neutron, charged particle and gamma photon activation is complicated by the fact that many elements produce more than one active nuclide in significant amounts. Direct resolution by least-squares fitting of the spectra of standards is only possible in these circumstances if the standard and sample spectra are obtained at the same time after irradiation, as the shapes of the standard spectra change with time. An alternative to the practical collection of spectra in this way is the correction of the standard spectra to the mid-time of counting of a sample spectrum. This may be achieved by recording spectra for a standard at different times and resolving the decay curves obtained for each channel on the basis of the half-lives of the component nuclides, which may be decay-independent or related or both. From the component nuclide count-rates in each channel at some arbitrary time, the standard spectrum at the time of counting of the sample can be generated and then used in a conventional least squares-fit of the sample spectrum. A FORTRAN IV program has been written to carry out this type of calculation on an IBM 360 65 computer. The feasibility of using this method is demonstrated by its application to activation analyses involving standards containing decay-related and independent nuclides.