The authors thank Professor Mei Jiang for her help in the statistical analysis and Mr. Hammer Tsui in the preparation of the manuscript. This study was supported by the Guangzhou Science and Technology Foundation (201804020043) and the National Natural Science Foundation of China (NSFC 81572063 and NSFC 81802076). The funding groups agreed with the study design, data analysis, manuscript preparation, and decision to publish. No other funding was received for this study.

This study and the use of human serum samples were approved by the Ethics Committee of The First Affiliated Hospital of Guangzhou Medical University (GYYY-2016-73). All participants have given their written consent independently or through their parents (in the case of children).
1. Basic information of the study group
NOTE: The Allergy Information Repository of the State Key Laboratory of Respiratory Disease (AIR-SKLRD) is a large serum bank established inside the Gu...

<ol>
	<li>Kamble, S., Bharmal, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incremental+Direct+Expenditure+of+Treating+Asthma+in+the+United+States.">Incremental Direct Expenditure of Treating Asthma in the United States.</a> Journal of Asthma Research. 46 (1), 73-80 (2009).</li><li>Paganelli, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+IgE+antibodies+in+the+diagnosis+of+atopic+disease.+Clinical+evaluation+of+a+new+in+vitro+test+system,+UniCAP,+in+six+European+allergy+clinics.">Specific IgE antibodies in the diagnosis of atopic disease. Clinical evaluation of a new in vitro test system, UniCAP, in six European allergy clinics.</a> Allergy. 53 (8), 763-768 (1998).</li><li>Wang, J., Godbold, J. H., Sampson, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+of+serum+allergy+(ige)+tests+performed+by+different+assay+systems.">Correlation of serum allergy (ige) tests performed by different assay systems.</a> Journal of Allergy and Clinical Immunology. 121 (5), 1219-1224 (2008).</li><li>Sun, B., Zheng, P., Wei, N., Huang, H., Zeng, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Co-sensitization+to+silkworm+moth+(Bombyx+mori)+and+9+inhalant+allergens+among+allergic+patients+in+Guangzhou,+Southern+China.">Co-sensitization to silkworm moth (Bombyx mori) and 9 inhalant allergens among allergic patients in Guangzhou, Southern China.</a> PLoS ONE. 9 (5), e94776 (2014).</li><li>Zeng, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Component-Resolved+Diagnostic+Study+of+Dermatophagoides+Pteronyssinus+Major+Allergen+Molecules+in+a+Southern+Chinese+Cohort.">Component-Resolved Diagnostic Study of Dermatophagoides Pteronyssinus Major Allergen Molecules in a Southern Chinese Cohort.</a> Journal of Investigational Allergology &amp; Clinical Immunology. 25 (5), 343-351 (2015).</li><li>Luo, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=grass+pollen+allergens+and+components+detected+in+a+southern+Chinese+cohort+of+patients+with+allergic+rhinitis+and/or+asthma.">grass pollen allergens and components detected in a southern Chinese cohort of patients with allergic rhinitis and/or asthma.</a> Molecular Immunology. 78 (2016), 105-112 (2016).</li><li>Zeng, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+profiles+of+serum+specific+IgE+and+IgG4+to+Dermatophagoides+pteronyssinus+allergen+and+its+major+components+during+allergen+immunotherapy+in+a+cohort+of+southern+Chinese+children.">Longitudinal profiles of serum specific IgE and IgG4 to Dermatophagoides pteronyssinus allergen and its major components during allergen immunotherapy in a cohort of southern Chinese children.</a> Molecular Immunology. 74 (2016), 1-9 (2016).</li><li>Lee, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+IgE+measurement+using+AdvanSure(R)+system:+comparison+of+detection+performance+with+ImmunoCAP(R)+system+in+Korean+allergy+patients.">Specific IgE measurement using AdvanSure(R) system: comparison of detection performance with ImmunoCAP(R) system in Korean allergy patients.</a> Clinica Chimica Acta. (9-10), 914-919 (2012).</li><li>Eckels, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+control,+analysis+and+secure+sharing+of+Luminex(R)+immunoassay+data+using+the+open+source+LabKey+Server+platform.">Quality control, analysis and secure sharing of Luminex(R) immunoassay data using the open source LabKey Server platform.</a> Bmc Bioinformatics. 14 (1), 145 (2013).</li><li>Bland, J. M., Altman, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Statistical+methods+for+assessing+agreement+between+two+methods+of+clinical+measurement.">Statistical methods for assessing agreement between two methods of clinical measurement.</a> Lancet. 327 (8476), 307-310 (1986).</li><li>Carletta, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+Agreement+on+Classification+Tasks:+The+Kappa+Statistic.">Assessing Agreement on Classification Tasks: The Kappa Statistic.</a> Computational Linguistics. 22 (2), 249-254 (1996).</li><li>Shyur, S. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+multiple+allergen-specific+IgE+by+microfluidic+immunoassay+cartridge+in+clinical+settings.">Determination of multiple allergen-specific IgE by microfluidic immunoassay cartridge in clinical settings.</a> Pediatric Allergy and Immunology. 21, 623-633 (2010).</li><li>Cesana, B. M., Antonelli, P., Gallazzi, E., Marino, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+measurement+methods:+an+endless+application+of+wrong+statistical+methods.">Comparison of measurement methods: an endless application of wrong statistical methods.</a> Intensive Care Medicine. 37 (6), 1038-1040 (2011).</li><li>Park, K. H., Lee, J., Sim, D. W., Lee, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+Singleplex+Specific+IgE+Detection+Immunoassays:+ImmunoCAP+Phadia+250+and+Immulite+2000+3gAllergy.">Comparison of Singleplex Specific IgE Detection Immunoassays: ImmunoCAP Phadia 250 and Immulite 2000 3gAllergy.</a> Annals of Laboratory Medicine. 38 (1), 23-31 (2018).</li><li>Teppo, H., Revonta, M., Haahtela, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allergic+rhinitis+and+asthma+have+generally+good+outcome+and+little+effect+on+quality+of+life+-+a+20-year+follow-up.">Allergic rhinitis and asthma have generally good outcome and little effect on quality of life - a 20-year follow-up.</a> Allergy. 66 (8), 1123-1125 (2011).</li><li>Fischer, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+of+type+I+sensitization+to+alpha-gal+in+forest+service+employees+and+hunters.">Prevalence of type I sensitization to alpha-gal in forest service employees and hunters.</a> Allergy. 72 (10), 1540-1547 (2017).</li><li>Li, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multicentre+study+assessing+the+prevalence+of+sensitizations+in+patients+with+asthma+and/or+rhinitis+in+China.">A multicentre study assessing the prevalence of sensitizations in patients with asthma and/or rhinitis in China.</a> Allergy. 64 (7), 1083-1092 (2009).</li><li>Ahlstedt, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+usefulness+of+specific+IgE+blood+tests+in+allergy.">Understanding the usefulness of specific IgE blood tests in allergy.</a> Clinical &amp; Experimental Allergy. 32 (1), 11-16 (2002).</li><li>Wood, R. A., Segall, N., Ahlstedt, S., Williams, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+IgE+antibody+laboratory+results.">Accuracy of IgE antibody laboratory results.</a> Annals of Allergy Asthma &amp; Immunology. 100 (2), 288-289 (2008).</li><li>Aberer, W., Kr&#228;nke, B., Hager, A., Wick, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+allergy+testing+needs+better+standardization--test+results+from+different+laboratories+lack+comparability+mostly+due+to+missing+effective+standards.">In vitro allergy testing needs better standardization--test results from different laboratories lack comparability mostly due to missing effective standards.</a> International Archives of Allergy &amp; Immunology. 108 (1), 82-88 (1995).</li></ol>

The authors have nothing to disclose.

Similar to the results from many other studies15,16,17, the results from the microfluidic system based on sera from 293 allergic patients showed that house dust mites (including D. pteronyssinus, D. farinae, and B. tropicalis) are the main inhalant allergens leading to allergic diseases in southern China, whereas for food, milk, peanut, shrimp, and crab are the commonest allergens that cause allergic s...

The prevalence of allergies has increased steadily in the past decades and is affecting 20%-30% of the global population1. Identification of the causative allergens has important significance in managing the diseases. In China, since registered in vivo skin prick tests are not available in the country, in vitro determination of serum-specific IgE is the most important and commonly used tool for type I allergy diagnoses2. This is similar to practices in the Western world, but although ImmunoCAP system (System 1), a fluorescence enzyme-linked immunosorbent base system, is perceived as the gold standard for in vitro allergy diagnosis3, its usage in China is very limited due to its high equipment and reagent price. Hence a new alternative allergy diagnosis system with a high price-performance ratio is badly needed.
The BioIC System (System 2) is a microfluidic cartridge-based system based on the chemiluminescence principle for multiplexed assays of serum-specific IgE. With a size of 7 cm x 4 cm, the microfluidic cartridge is composed of three layers of injection-molded plastic. The upper part is 3 mm-thick transparent polycarbonate which carries good stability during thermal assembly processes. Together with the 3 mm-thick bottom layer constructed from a copolymer of acrylonitrile, butadiene, and styrene (ABS), it sandwiches the 0.5 mm-thick middle layer made of silicone rubber. Being black in color, the middle layer offers lower background during chemiluminescence detection. On top of the silica gel, a thin layer of nitrocellulose membrane (NC membrane) is sprayed at the position corresponding to the reaction zone, which allows the spotting of different allergenic proteins. The purpose of this study is to evaluate the clinical performance of the microfluidic system for the multiplexed determination of allergen-specific IgE in serum.

<table>
	<tbody>
		<tr>
			<td>Agnitio BioIC Analyzer</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>BA-G2000</td>
			<td></td>
		</tr>
		<tr>
			<td>BioIC Allergen specific-IgE Detection Kit</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>DR17A12</td>
			<td></td>
		</tr>
		<tr>
			<td>BioIC Cartrideg Placement plate</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>T20SET</td>
			<td></td>
		</tr>
		<tr>
			<td>BioIC Reagent Dispenser</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>DS-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Image two-dimensional barcode machine</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>NLS-HR200</td>
			<td></td>
		</tr>
		<tr>
			<td>Software Package, LabIT</td>
			<td>Agnitio Science &#38; Technology(Taiwan, China)</td>
			<td>Version 2.4.12</td>
			<td></td>
		</tr>
		<tr>
			<td>VORTEX-5 Vortex Mixer</td>
			<td>Haimen Kylinbell Lab Lastruments Co., Ltd.</td>
			<td>VORTEX-5</td>
			<td></td>
		</tr>
	</tbody>
</table>

application of biochip microfluidic technology to detect serum allergen-specific immunoglobulin e (sige)

Allergic disease is common in both adults and children. Identification of the causative allergens is significant in disease management and prevention. However, a specific immunoglobulin E (IgE) measurement system with a high price-performance ratio is lacking in mainland China, especially in the primary care hospitals. This paper describes the principle and operation procedures of using a microfluidic cartridge-based chemiluminescence system to detect allergen-specific IgE in serum. The results were compared with those from ImmunoCAP (System 1), the industrial standard, and the reproducibility of the system to detect patients sensitized to common allergens is evaluated. The results showed that in comparison with ImmunoCAP (System 1), the BioIC System (System 2) has good precision and sensitivity in detecting serum-specific IgE against various inhalant and food allergens but with a significantly lower cost. It can serve as a good alternative to System 1 in primary care hospitals in mainland China who have lower financial affordability.

Positive rates for 19 common allergens 
The results on 293 sera are shown in Figure 3. Among all the inhalant allergens, D. farinae had the highest positive rate (80.89%, 273/293), followed by D. pteronyssinus (78.84%, 231/293). Among the food allergens, crab has the highest positive rate (20.48%, 60/293), followed by 13.65% (40/293) for shrimp. The total positive rate for inhalant allergens was higher than tha...

Watch this Scientific Journal Video about Application of Biochip Microfluidic Technology to Detect Serum Allergen-specific Immunoglobulin E sIgE at JoVE.com

Application of Biochip Microfluidic Technology to Detect Serum Allergen-specific Immunoglobulin E sIgE

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.

Application of Biochip Microfluidic Technology to Detect Serum Allergen-specific Immunoglobulin E (sIgE)

Allergic disease is common in both adults and children. Identification of the causative allergens is significant in disease management and prevention. ...

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.

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16.2K 조회수.  First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University. 바이오칩 미세 유체 기술 시스템은 혈청 의 염기 IGE에 대한 화학 발광 원리를 사용하는 미세 유체 카트리지 기반 섹션입니다. 다른 검출 방법에 비해, 바이오IC 시스템은 저렴한 비용과 혈청 IGE를 감지하기 위해 적은 혈청을 필요로한다는 장점이 있습니다. 이 기술의 의미는 비염과 천식과 같은 알레르기 질환의 진단을 쪽으로 경향이 있다 이 환자의 많은 일반적인 검출 가능?...