Name: evaluation of a point-of-care testing analyzer for measuring peripheral blood leukocytes
Uploaded: 2022-03-22T15:00:00
Description: Watch this Scientific Journal Video about Evaluation of a Point-of-Care Testing Analyzer for Measuring Peripheral Blood Leukocytes at JoVE.com

This study was supported by the Medical Scientific Research Foundation of Guangdong Province, China (A2019224). 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 blood 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: Venous blood was collected from patients who visited the First Affiliated Hospital of Guangzhou Medical University (Hospital 1) ...

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With the advancement of modern laboratory medicine, it is now typical to see several detection technologies utilized in the same or different laboratories to identify the same clinical marker. As a result, more emphasis should be placed on the consistency of test results to assist clinics in making accurate interpretations and judgments of test results. According to the investigation, the total value of laboratory equipment in tertiary hospitals and independent laboratories is substantially higher than that in primary ho...

White blood cell (WBC)count or differential is an important indicator to reflect the inflammation of the body, which can distinguish bacterial infection from viral infection. WBC analysis is also helpful to guide the follow-up diagnosis and treatment1. At present, the five-classification fully automatic hematology analyzer has been widely used in large and medium-sized medical units, because it is automatic, has high efficiency, yields accurate and reliable results, and effectively reduces the work intensity of laboratory technicians. It plays an important role in clinical examination2,3. However, most primary medical institutions, such as community healthcare centers and private clinics, have a low adoption rate of a hematology analyzer. According to a nationwide multicenter study on clinical laboratory construction in China, the laboratory construction of primary medical institutions is insufficient, as evidenced by the small size of laboratories, the insufficient talents transmission, and the spread of science and technology to the countryside, amongst other factors4.
Since December 2019, COVID-19 began to spread all over the world and developed into a global pandemic. In the 'post-epidemic era', a series of national policies have been proposed to implement the normalized prevention and control measures of epidemic situations. The laboratory of primary medical institutions plays an important role in grassroots diagnosis and treatment and disease prevention and control. It is the first line of defense and control in epidemic situations, and it is critical to COVID-19 prevention and control5. Some studies have shown that the detection of peripheral blood lymphocytes and neutrophils will contribute to COVID-19 patient screening, diagnosis, and treatment, and that the neutrophil/lymphocyte ratio can also be used as clinical early warning indicators of severe and critical COVID-196,7. Moreover, leukocyte detection has the benefit of providing a quick report. Primary medical and health institutions can extensively carry out leukocyte detection to help detect and screen suspected infections in time.
POCT card-based leukocyte analyzer (evaluated system; see Table of Materials) is a three-classification blood cell analyzer based on the gold standard "Coulter principle". The evaluated system provides quantitative analysis results of one WBC histogram and seven blood parameters including WBC count, number of granulocytes (Gran#), percentage of granulocytes (Gran%), number of lymphocytes (Lym#), percentage of lymphocytes (Lym%), number of intermediate cells (Mid#), and percentage of the intermediate cells (Mid%). It adopts the card-based innovative technology and has advantages such as the availability of single-person detection kit, absence of liquid waste, fast detection in 30 s, being free from routine maintenance, and user-friendly operation. Therefore, it is particularly well-suited to primary medical institutions. This study aims to evaluate the clinical detection performance of POCT card-based leukocyte analyzer by comparing against two fully automatic commercial hematology analyzers (reference system 1 and reference system 2; see Table of Materials) from laboratories of two large-scale public hospitals.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Blood cell detection module</td>
			<td>Chuanghuai Medical Technology Co., Ltd.(Shenzhen, China)</td>
			<td></td>
			<td>consumables for evaluated system</td>
		</tr>
		<tr>
			<td>Blood lancet</td>
			<td>Chuanghuai Medical Technology Co., Ltd.(Shenzhen, China)</td>
			<td></td>
			<td>consumables for evaluated system</td>
		</tr>
		<tr>
			<td>Hemolytic reagent</td>
			<td>Chuanghuai Medical Technology Co., Ltd.(Shenzhen, China)</td>
			<td></td>
			<td>consumables for evaluated system</td>
		</tr>
		<tr>
			<td>IBM SPSS Statistics 25</td>
			<td>International Business Machines Corp., Armonk, NY</td>
			<td></td>
			<td>Software for data analysis</td>
		</tr>
		<tr>
			<td>MedCalc 11.4.2.0</td>
			<td>2021 MedCalc Software Ltd</td>
			<td></td>
			<td>Software for data analysis</td>
		</tr>
		<tr>
			<td>Microsoft Excel 2019</td>
			<td>Microsoft</td>
			<td></td>
			<td>Software for data analysis</td>
		</tr>
		<tr>
			<td>Point-of-care testing (POCT) card-based leukocyte analyzer</td>
			<td>Chuanghuai Medical Technology Co., Ltd.(Shenzhen, China)</td>
			<td>CX-2000</td>
			<td>Evaluated system</td>
		</tr>
		<tr>
			<td>Quantitative pipette with capillary tube inside</td>
			<td>Chuanghuai Medical Technology Co., Ltd.(Shenzhen, China)</td>
			<td></td>
			<td>consumables for evaluated system</td>
		</tr>
		<tr>
			<td>Siemens fully automatic hematology analyzer and its related reagents and consumables</td>
			<td>Siemens Healthcare Diagnostics Inc.</td>
			<td>ADVIA 2120i</td>
			<td>Reference system 2</td>
		</tr>
		<tr>
			<td>UniCel DxH 800 Coulter Cellular Analysis System and its related reagents and consumables</td>
			<td>Beckman Coulter, Inc.</td>
			<td>DxH 800</td>
			<td>Reference system 1</td>
		</tr>
	</tbody>
</table>

evaluation of a point-of-care testing analyzer for measuring peripheral blood leukocytes

White blood cell (WBC) is an important indicator of inflammation in the body, and it can help distinguish between bacterial and viral infections. At present, most primary medical institutions in China have a poor percentage of adoption of blood-testing technology, and a hematology detection system with a high price to performance ratio and easy operation is urgently needed in primary healthcare centers. This paper introduces the principle and operation procedures of a point-of-care testing (POCT) card-based leukocyte analyzer (evaluated system), which was used to detect WBC indexes such as neutrophils, lymphocytes, and intermediate group cells (including eosinophils, basophils, and monocytes) in whole blood. The results from the evaluated system were compared to those from two commercial automatic hematology analyzers (reference system). The correlation and consistency between the evaluated system and the commercial reference systems were analyzed. The results showed that WBC count and number of granulocytes detected by the evaluated and reference systems showed a strong positive correlation (rs = 0.972 and 0.973, respectively), while the number of lymphocytes showed a relatively low correlation (rs = 0.851). A Bland-Altman plot showed that the major difference between the values detected by the evaluated system and the reference systems is within 95% limits of agreement (LoA), indicating that the two systems are in good agreement. In conclusion, the evaluated system has an excellent correlation, robust consistency, and a reliable comparison with the results of the widely used automatic hematology analyzers. It is ideal for WBC detection in primary medical institutions where a full-automatic five-category hematology analyzer is unavailable, especially during the COVID-19 normalized prevention and control period.

Sample data 
A total of 1066 patients were enrolled in two research centers, including Hospital 1 (n = 532) and Hospital 2 (n = 534). The patient characteristics are shown in Table 1. The percentage of males is 49.9% and the median age is 52 (32, 66) years. Patients enrolled in the study were comprised of inpatients (51.1%), outpatients (39.0%), and physical examination patients (8.4%). The samples tested were from patients who visited the internal medicine departments (30.6%), surg...

Watch this Scientific Journal Video about Evaluation of a Point-of-Care Testing Analyzer for Measuring Peripheral Blood Leukocytes at JoVE.com

Evaluation of a Point-of-Care Testing Analyzer for Measuring Peripheral Blood Leukocytes

A protocol to measure peripheral blood leukocytes using a POCT card-based leukocyte analyzer is presented here. Same blood samples were tested by two automated hematology analyzers to evaluate the consistency and accuracy of the results. The results showed that the evaluated analyzer had a good correlation with the reference system.

White blood cell (WBC) is an important indicator of inflammation in the body, and it can help distinguish between bacterial and viral infections. At ...

A detailed description of the standardized operation of the analyzer in the protocol can help a person who has never operated the analyzer to complete the experiments mostly important, accurate and with reliable results. Using Jove's innovative technology has advantages such as the availability of a single person detection kit, absence of liquid waste, being fast, being free for routine maintenance, and user friendly operation. One should first understand the detection principle of the analyzer, and then be familiar with several critical steps in the protocol such as blood collection, reagents lifting and so on.To begin start the analyzer by turning the off or in power switch on the back of the analyzer to in and ensure that the indicator light of the analyzer is on. Enter username and password in the login dialogue box and click on login. The system should perform self check and start initialization automatically and display the sample analysis page.Start the test preparation by clicking on next sample then enter the gender, name and other clinical information correctly and select the appropriate reference group. Later, click on okay to save the information. Tear the thin film of the blood cell detection module, press the entry exit warehouse button and place the blood cell detection module into the machine warehouse correctly, with its orifice facing outward.Puncture the hemolytic reagent ceiling film with the tip of a quantitative pipette. For capillary blood collection, disinfect the left ring finger once with a cotton swab dipped in alcohol one way. After the alcohol naturally dries out, use a blood lancet to puncture the skin of the left ring finger.Gently squeeze out the first drop of blood and wipe it with a cotton swab. Squeeze out enough blood to form a full water drop and collect five microliters of the blood sample using the capillary tube inside the quantitative pipette. For reagent mixing, insert the quantitative pipette into the hemolytic reagent and press it tightly to release the blood sample from the capillary tube.Mix the blood in the capillary tube and hemolytic reagent by turning it upside down 15 to 20 times at a constant speed until no red blood remains in the capillary tube. Start sample analysis by opening the lid and squeezing the solution into the blood cell detection module. Next, press the entry exit warehouse button.After the blood cell detection module enters the warehouse press the counting button. On the analyzer interface click on the okay button twice to confirm that the blood cell detection module has been taken out. On the analyzer interface click on the print button to print the test results.To turn off the analyzer, click on the shutdown button on the analyzer interface and select yes in the dialogue box that pops up on the interface. Check that the system starts to execute the shutdown sequence. Set the off or in switch on the back of the mainframe to off after the shutdown sequence is completed.The figure shows the scatter plot and Spearman correlation analysis of the evaluated and reference systems. The white blood cell count and granulocyte number exhibit the strongest correlation. Whereas, the lymphocyte number exhibits the weakest correlation between the evaluated and reference systems.A Bland-Altman plot was obtained to visualize the consistency analysis. The white blood cell results detected by the evaluated system agree with the reference system. The average values of the differences in the leukocyte indexes are above zero, indicating that the test results of the evaluated system are slightly higher than those of the reference system.During the experiment we should try our best to ensure that the blood warrant we collected is exactly 5 microliter and the in room side are completely dissolved by the hemolytic reagent.

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