TITLE: No Title
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[DESCRIPTION]
<p>The appropriate use of clinically accurate diagnostic tests is essential for the detection of pertussis, a poorly controlled vaccine-preventable disease. The purpose of this study was to estimate the sensitivity and specificity of different diagnostic criteria including culture, multi-target polymerase chain reaction (PCR), anti-pertussis toxin IgG (IgG-PT) serology, and the use of a clinical case definition.  An additional objective was to describe the optimal timing of specimen collection for the various tests. </p>
<p>Clinical specimens were collected from patients with cough illness at seven locations across the United States between 2007 and 2011. Nasopharyngeal and blood specimens were collected from each patient during the enrollment visit.  Patients who had been coughing for ≤ 2 weeks were asked to return in 2-4 weeks for collection of a second, convalescent blood specimen.  Sensitivity and specificity of each diagnostic test were estimated using three methods—pertussis culture as the “gold standard,” composite reference standard analysis (CRS), and latent class analysis (LCA). </p>
<p>Overall, 868 patients were enrolled and 13.6% were <em>B. pertussis</em> positive by at least one diagnostic test.  In a sample of 545 participants with non-missing data on all four diagnostic criteria, culture was 64.0% sensitive, PCR was 90.6% sensitive, and both were 100% specific by LCA.  CRS and LCA methods increased the sensitivity estimates for convalescent serology and the clinical case definition over the culture-based estimates.  Culture and PCR were most sensitive when performed during the first two weeks of cough; serology was optimally sensitive after the second week of cough.</p>
<p>Timing of specimen collection in relation to onset of illness should be considered when ordering diagnostic tests for pertussis. Consideration should be given to including IgG-PT serology as a confirmatory test in the Council of State and Territorial Epidemiologists (CSTE) case definition for pertussis.</p>

[BEFORE_START]
<ul>
<li><strong>Disclaimer:</strong> Names of vendors or manufacturers are provided as examples of suitable product sources; inclusion does not imply endorsement by the Centers for Disease Control and Prevention or the Department of Health and Human Services.</li>
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<p><strong> </strong></p>
<p>Reagents:  96 well microtiter strip assembly; Pertussis toxin (PT) antigen, in 50% (v/v) glycerol; Coating buffer, (0.05 M Carbonate – bicarbonate buffer, pH 9.6); IgG anti-PT standards (Standard) (lyophilized) A – F (15, 30, 60, 120, 240 &amp; 480 International Units (IU)/mL); IgG anti-PT controls I (negative), II (26-68 IU/mL) &amp; III (56-134 IU/mL) (Kit controls) (lyophilized); Assay buffer, (Phosphate buffer saline containing 4% BSA &amp; 0.05% Tween – 20); Standard &amp; Kit control diluent (Deionized water containing 0.05% Tween–20); 10X Wash solution concentrate, (10X PBS containing 0.5% Tween–20); Peroxidase labeled mouse monoclonal antihuman IgG conjugate (Conjugate) (Provided as a 1:5,000 dilution in 50% glycerol); TMB substrate, (3,3’,5,5’ tetramethylbenzidine in mildly acidic buffer); and, Stop solution, (1 N Hydrochloric acid).</p>
<p> </p>
<p>Equipment/Materials: Adhesive microtiter plate / strip sealers (plate sealer); Disposable reagent reservoirs; Decapper; Deionized water;  Microtiter plate or strip washer; Microtiter reader equipped to read absorbance at 450 nm; Software with the capability to use the four-parameter logistic (4PL) function for the analysis of absorbance data from a 96-well plate layout; Single and multichannel micropipettes with appropriate tips; Repeater pipettes with appropriate tips; Class A glassware; graduated cylinders, and beakers; Opaque box, slightly larger than microtiter plate / strip assembly; Thermometer; Timer; Vortex mixer; 50mL Conical tubes; 5mL Test tubes; and, Reservoir basins.</p>

[GUIDELINES]
<p>Title: Standard Operating Procedure for the Detection of Human, IgG Anti-Pertussis Toxin.</p>
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<p>Purpose: To describe the procedures used to estimate IgG antibodies against Pertussis Toxin (PT) in human serum by the Human, IgG Anti-Pertussis Toxin ELISA<em>.</em></p>
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