This paper presents a protocol for detecting serum-specific immunoglobulin E with a microfluidic cartridge-based chemiluminescence system and the evaluation of its usefulness in allergy diagnoses.
The biochip microfluidic technology system is a microfluidic cartridge based section that uses the chemiluminescence principle for assage of serum specif IGE. Compared with other detection methods, the bioIC systems has the advantages of being low cost and requiring less serum to detect serum IGE. The implications of this technique is tend towards the diagnosis of allergy diseases such as rhinitis and asthma because many of these patients are sensitized to common detectable allergens.
Although this method can provide insight into respiratory allergy diseases, it can also be applied to other situations, such as food and skin allergies. As this technique is easier to use, and has a lowered method for operator training, individuals new to the system can learn to use it quickly. Vivid demonstration of this matter is critical as the determines whether the test is successful and requires skillful manual operation.
To test serum samples for serum immunoglobulin E antibodies to whole allergens, randomly select three samples with at least 900 microliters of serum and take the chip from the kit and number it. Then, turn on the computer and the analyzer power. Start the Lab IT program on the computer.
If the dark frame warning window pops up, click OK to run a leak test, then click the center logo to enter the operation interface. Check the reaction temperature and the charge coupled device or CCD temperature, the reaction temperature should rise to 37 plus or minus one degree celsius in about ten minutes and the CCD temperature should drop to minus 15 plus or minus one degree celsius. When the CCD temperature has dropped to minus 15 plus or minus one degree celsius, confirm that there are no other items left inside the instrument, and close the door.
Then click tools, system test, leak test. When the test is finished, the report window will pop up. Next, load 620 microliters of wash buffer, 120 microliters of blocking buffer, 60 microliters of conjugates A and B, 60 microliters of substrates A and B, and 100 microliters of each serum sample into the corresponding reagent tanks on the microfluidic cartridge.
After clicking cartridge ID, use the bar code scanner to scan the serial number of the cartridge and enter the sample ID.Then place the cartridge into the analyzer, close the door, and click analyzer and run to start the analysis. At the end of the analysis, export the results into an appropriate statistical software program and remove the cartridge from the analyzer. Lightly wipe the internal heating iron and electromagnet of the analyzer with 75 percent alcohol and close the Lab IT window.
The temperature monitoring window will pop up and will automatically close when the CCD warms up to the five degree celsius protection mode, at which point it is safe to turn off the analyzer and the computer. The microfluidic system produces repeatable results, as demonstrated by these data for cat dander, dog dander, and cockroach after nine rounds of testing. As observed, all of the data are within the range of two standard deviations, which is consistent with the maximum allowable clinical error.
As expected from the positivity rates for the allergens, all of the evaluation results for inhalant allergens are superior to the food allergens. Since the area under the curve values demonstrate a range from 0.613 to 0.984, and the area under the curve values for the three inhalant allergens are greater than 0.950, it can be concluded that system two has a high accuracy with reference to system one. Consistency analysis for the two systems reveals that the kappa values for the three inhalants are all greater than the kappa values for the food allergens which in general fall below 0.4.
In these scatter plots constructed with system two results along the x-axis and system one results along the y-axis, the distribution of the serum immunoglobulin E concentration results from the two systems for six different allergens can be observed. Before starting the detection, always check the reaction and CCD temps as they are linked to the accuracy of the test. Following this procedure, other methods like component resolved diagnosis can be performed to answer additional questions about allergen cross standardization and co-sensitization.
Microfluidic chip technology has been successfully applied in the field of allergy diagnosis to address problems such as inadequate serum, and high medical cost As working with human serum, symptoms can be determining hazardous. Wearing the appropriate personal protection equipment and other precautions should always be taken while performing this procedure.
