The overall goal of this experiment is to investigate the sensitization to various grass pollen allergen components in southern China. This med can help answer key questions in the energy field about how individuals become sensitized to grass pollens. Main advantage of this technique is that the use of recombinant or purify allergen molecules.
Can also convey the occurrence of confounding rashes, providing a better test accuracy. The implication of this technique is then to thwart the diagnosis of allergy, rhinitis, and asthma. As many of these patients are sensitized to common detectable allergens.
Though this method can provide insight into allergy airway disease, it can also be a to other type of allergies such as food allergies. We first had the idea for this method when you found or CID for Chinese Demonstrating the procedure will be Nili Wei, a from my lab. After retrieving the serum samples of eligible patients from the biobank.
Turn on the fully automated immunoassay analyzer and the built in information data management computer. With the primary power on, switch the instrument to the system power and wait three minutes for the built in software to start. Then add 140 microliters of serum to one vile per allergen and or allergen component test.
And label each vile of serum with an identification number unique to each patient. From the information data management interface, open the request list window to confirm that serum IGE is the test method of choice. Then load the sample tubes into the sample racks.
And the barcode labeled quality control tubes into the quality control racks. In the assay processing menu, select, load reagents to complete the loading of the sample and quality control racks, development solution, conjugate, calibraters, carrier, pipette tips, stop solution, and washing solution, according to the load list. Click load rinse solution and load washing solution.
When all of the materials have been loaded select load and start and click okay. At the end of the measurement, the results will appear in the information data management interface. Select the data to be analyzed and click menu, approve, and save as to export the serum IGE measurement results as a spreadsheet file.
Then import the spreadsheet file into an appropriate statistical software program. And use a specific module for Spearman's coefficient to analyze the correlations in sensitization between the grass pollens and the pollen components. Of the 258 respiratory allergy patients studied in this experiment, 22.5 percent tested positive to Bermuda grass, 13.6 percent to Timothy grass, and seven percent to Hummulus scandens.
Interestingly, patients with allergic rhinits and or asthma demonstrated a remarkably higher rate of sensitization to Bermuda grass than did patients with other allergies. Among the patients with Bermuda grass class one serum IGE reactivity, the likelihood of positive tests for Cyn d 1, Timothy grass, or Humulus scandens allergens or cross reactive carbohydrate determinants was below 30 percent. In contrast, patients with class four Bermuda grass SIGE reactivity exhibited a 67 to 100 percent chance of concomitant positivity to these grass components.
Among the 35 Timothy grass positive patients, the most frequently detected allergen component was Phl p four followed by Phl p one. Three patients each were positive for any one of Phl p five, six, seven, 11, or 12. Serum Phl P four SIGE levels correlated strongly with total Timothy grass allergen and cross reactive carbohydrate determinants SIGE levels.
41.4 percent of Bermuda grass positive patients and 68.6 percent of Timothy grass sensitized patients were also SIGE positive to cross reactive carbohydrate determinants. With Spearman's rank correlation analyses demonstrating a significant association in sensitization between cross reactive carbohydrate determinants and other cross allergens. Following this procedure are the methods like the of immunoassay analyzers can be performed to answer additional questions about sensitization to grass pollen allergy in the center or nation-wide study.
While attending this procedure, it's important to remember to insure the appropriate and correct calibration has been growed in the system. And this is essential for accurate test results. After watching this video, you should have a good understanding of how to investigate patient sensitization to components of grass pollen allergies.
Don't forget that working with human serum samples can be extremely dangerous. And appropriate precautions against pathogen infections should always be taken while performing this procedure.
