Immunofluorescence Lateral Chromatography in Combination

Materials Express

Immunoﬂuorescence lateral chromatography in combination with detection of hypersensitive C-reactive protein and procalcitonin levels in patients with mental disease associated with coronavirus 2019 disease infection

Dan Li1,†, Xiaoliang Zhou2,†,YunShi1,†,PingLi1, Shuzi Chen1, Weifeng Jin1, Xiaoyan Lou1, Zhenhua Li1, Ping Lin1,∗, and Yang Sun3,4,∗ 1Department of Clinical Laboratory, Shanghai Mental Health Center, Shanghai 200030, PR China 2Department of Clinical Laboratory, Wuhan Mental Health Center, Wuhan 430022, Hubei, PR China 3Institute of Arthritis Research, Shanghai Academy of Chinese Medical Sciences, Shanghai 200052, PR China 4Guanghua Integrative Medicine Hospital, Shanghai 200052, PR China IP: 192.168.39.210 On: Sun, 26 Sep 2021 07:58:46 Copyright: American Scientific Publishers Delivered by Ingenta

ABSTRACT Article Immunofluorescence lateral chromatography (IFLC) is a quantitative detection technique used to screen C-reactive protein and procalcitonin (PCT) monoclonal antibodies with high efficiency and specificity. This study investigated the application value of IFLC combined with the detection of hypersensitive C-reactive protein (hsCRP) and PCT in patients with mental diseases associated with COVID-19 infection. Sixty-four patients with mental disease admitted to our hospital from January to April 2020, were selected as research subjects. Based on the diagnostic criteria for COVID-19 in the “Corona Virus Disease 2019 Diagnosis and Treatment Scheme (Byass, P., 2020. Eco-epidemiological assessment of the COVID-19 epidemic in China, January- February 2020. Global Health Action, 13(1), p.1760490.) version 6, all patients were tested using the nucleic acid amplification test for COVID-19 and were divided into two groups of 6 infected and 58 non-infected patients. Subsequently, the patients’ hsCRP and PCT levels were measured using IFLC and an automatic biochemistry analyzer (BA). The results showed that the hsCRP and PCT levels in the infected group were higher than the non-infected group. Additionally, the analytical ultracentrifugation (AUC), sensitivity, and specificity of hsCRP were higher than PCT. The AUC, sensitivity, and specificity of hsCRP+PCT measured jointly were significantly higher than hsCRP and PCT measured separately. The BA results showed that the AUC, sensitivity, and specificity of hsCRP were higher than PCT. The AUC, sensitivity, and specificity of hsCRP + PCT measured jointly were significantly higher than hsCRP and PCT measured separately. Comprehensive analysis showed that the AUC, sensitivity, and specificity of hsCRP+PCT tested using IFLC were significantly higher than using the BA. All of the above differences were statistically significant (P<0.05). It was concluded that the joint detection of hsCRP and PCT by IFLC helped identify and diagnose mental diseases associated with COVID-19 infection.

∗Authors to whom correspondence should be addressed. †These three authors contributed equally to this work.

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Keywords: Immunoﬂuorescence, Joint Detection, COVID-19, Hypersensitive C-Reactive Protein, Procalcitonin.

1. INTRODUCTION 2. MATERIALS AND METHODS When the blood or respiratory tract samples tested using 2.1. Materials real-time fluorescence quantitative polymerase chain reac- 2.1.1. Patient Information tion are positive for the novel Coronavirus 2019, or The patients with mental diseases admitted to the Wuhan their viral gene sequencing is significantly homologous Mental Health Center, Wuhan, China, from January to to it, patients are confirmed to be infected with COVID- April 2020, were selected as the research subjects. Inclu- 19 (Coronavirus 2019 Disease) [1]. Mental illness is a sion criteria: (1) meet the diagnostic criteria for mental ill- special kind of disease. A patients’ mental state may ness [5]; (2) the age of 18 years old or more; (3) no recent be affected by their perception of COVID-19 symptoms history of surgery or trauma; (4) family members sign that could even lead to the aggravation of hypochondria, informed consent forms. Exclusion criteria: (1) concomi- social phobia, generalized anxiety disorder, and obsessive- tant immune system diseases; (2) combined with infec- compulsive disorder, which is not conducive to the treat- tious diseases other than COVID-19; (3) complicated with ment of mental illness [2]. COVID-19 is characterized malignant tumor; (4) suffering from heart, liver, kidney by fever, dry cough, fatigue, and other clinical manifesta- and other important visceral lesions. Sixty-four patients tions. It is highly contagious and insidious and can occur were enrolled in the study. Based on the diagnostic criteria in people of all ages. Currently, the nucleic acid test is for COVID-19 in the “Corona Virus Disease 2019 Diag- frequently used in the clinical diagnosis of COVID-19, nosis and Treatment Scheme” version 6, all patients were but it is significantly affected by external factors, lead- tested using the nucleic acid amplification test [6]. They ing to some false-negative results, and there are defi- were divided into an infected group (n = 6) and a non- ciencies in sensitivity and specificity [3]. Therefore, it is infected group (n = 58). 2 detect was used to compare the Article important to find an effective, rapid, and accurate method genders of the two groups, while t test was used to com- for identifying COVID-19 infected patients so that they IP: 192.168.39.210 On: Sun,pare 26 theSep age, 2021 height 07:58:46 and body mass index, and the differ- may obtain timely and effective treatmentCopyright: and stopAmerican the Scientific Publishers ences were not statistically significant (P>0.05) (Table I). spread of the virus. The quantitative detection techniqueDelivered by Ingenta This study was approved by the Medical Ethics Committee of immunofluorescence lateral chromatography (IFLC) is of our hospital. based on the reaction principle of the double-antibody sandwich system. The technique selects monoclonal anti- 2.1.2. Reagents and Suppliers bodies against highly specific target proteins to pair with each other and then produces fluorescence-labeled conju- All reagents were used in this study were analytically · gation; the cloaked, captured antibodies form a double- pure: Na2HPO4 12H2O (disodium hydrogen phosphate) antibody sandwich system, and finally, it detects the target (article No.: 0404, Shanghai Liansuo Biotechnology Co., proteins in serum sample [4]. HsCRP is an acute phase Ltd.), triton-x100 (article No.: X-100, Haian Petrochemi- protein synthesized by liver cells; it can increase sharply cal Plant, Jiangsu Province, China), tris (article No.: 0161 when the body is infected and returns to normal range on AMRESCO), tween-20 (article No.: P137, Shanghai Lian- the remission of the disease and recovery of tissues and suo Biological Technology Co., Ltd.), 99% PROCLIN- structural functions. Therefore, the level of hsCRP reflects 300 (article No.: ZB116, Shanghai Jinjing Biotechnology the occurrence, development, and prognosis of infectious Co., Ltd.), 99% trehalose (article No.: YY11150, Shanghai diseases. PCT is a protein that increases with protozoan, Yuanye Biotechnology Co., Ltd.) polyethylene glycol 600 systemic bacterial or other bacterial infections. Therefore, (article No.: 167161, Xibao Biological Technology, Co., the occurrence and development of infectious diseases can Ltd., Shanghai), fluorescent microspheres F1-XC005 and be accompanied by in vivo changes in serum hsCRP and F1-XC03 (Hongrong Weizre Bioengineering Technology PCT levels. In this study, patients with mental illness and Co., Ltd., Shanghai), polyester fiber film, cellulose nitrate infected with COVID-19 were the research subjects, while film CN95, and glass fiber film 8964 (Xuxingtai Quan- non-infected patients with mental illness were the controls. tity Technology Co., Ltd.) PVP K30 (polyvinylpyrroli- In addition to IFLC, an automatic biochemistry analyzer done) (article No.: J0029-1, Suzhou Yaco Technology Co., (BA), commonly used in clinical practice, was used for Ltd.) and PVA (polyvinyl alcohol), article No.: S25454 testing study participants. The purpose of the study was (Shanghai Yuanye Biotechnology Co., Ltd.), CRP mon- to explore the effect of IFLC combined with hsCRP and oclonal antibodies C2, C6, PCT monoclonal antibodies PCT detection on the diagnosis of COVID-19 and provide 42, and 16B5 (Shanghai Yibo Biotechnology Co., Ltd.), a reference for the clinical optimization of the diagnostic CM-EUs (Thermo Fisher Scientific Inc.), sample pads and process. absorbent pads (Bedford, Massachusetts, USA).

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Table I. Comparison of data between the infected and non-infected groups.

Gender [case, (n)%] Body mass index Group Male Female Age (x ± s, year) Height (x ± s,cm) (x ± s, kg/m2)

Infected group (n = 6) 4 (66.67) 2 (33.33) 45.29 ± 6.54 163.03 ± 5.02 23.05 ± 3.31 Aon-infected group (n = 58) 28 (48.28) 30 (51.72) 45.37 ± 6.47 162.89 ± 5.04 22.94 ± 3.58 2/t 0.184 0.029 0.065 0.072 P 0.668 0.977 0.949 0.943

2.2. Methods microsphere [7, 8]. After activating the fluorescent micro- 2.2.1. Quantitative Immunofluorescence Lateral spheres, the coupled buffer (0.02 M PB, pH 7.4) was taken Chromatography to wash the activated fluorescent microspheres twice ultra- The flowchart for the method is shown in Figure 1. The sonically. Then, the CRP antibody was added as required hsCRP and PCT levels of all patients were determined by to obtain the fluorescence-labeled conjugation (concentra- IFLP. The steps are as follows: the sample diluent was tion: 0.2–1 mg/mL), and the reaction container was oscil- prepared (PEG 6000: triton-X100: proclin-300: buffer = lated at 37 C for 120–180 min. The prepared coupling 0.5%:0.5%:0.02%:0.05 M PBS (Phosphate Buffer Saline), buffer was used to change the liquid and then oscillated pH 7.4), sodium phosphate buffer (0.01 M PBS, pH 7.4), continuously at 37 C for 30 min. After centrifugation, tris-Cl buffer solution (0.1 M pH 8.0), tris-Cl buffer solu- the supernatant was discarded. Storage buffer of the same tion (0.01 M, pH 8.0), tris-Cl buffer solution (pH 8.0), volume was taken for ultrasonic washing, re-suspended, activation buffer (0.01 M NaH2PO4, pH 6.0), coupling precipitated, re-dissolved, and stored at 4 C in the dark. buffer (0.02 M PB (Phosphate Buffer), pH 7.4), and stor- (2) Fluorescence-labeling, conjugation and curing. Pre- age buffer for standby use. treatment buffer solution with added PVP, PVA, tween- The europium (III) chelate microparticle (CM-EUS) test 20, triton X-100, and BSA was prepared to dilute the strip comprises a sample pad, a markerIP: 192.168.39.210 pad, a coated mem- On: Sun, 26 Sep 2021 07:58:46 brane, and an absorption pad. (1) FluorescentCopyright: microsphere American Scientific Publishers conjugated antibody. The coupling principle ofDelivered carboxyl by Ingenta modified fluorescent microspheres and antibodies is shown in Figure 2. CRP monoclonal antibody C6 was selected, Article and the conjugated ratio was 0.5 mg antibody: 10 mg

Fig. 2. Coupling principle of carboxyl modiﬁed ﬂuorescent microspheres and antibodies. Under weakly acidic conditions, microspheres were activated by soluble EDAC and N -hydroxysuccinimide to form intermediates, prompting their surface –COOH to covalently bond with

Fig. 1. Flowchart of the two detection methods. QDTOILC: Quanti- –NH2 in the Fc region of IgG molecules for the coupling of ﬂuorescent tative detection technique of immunoﬂuorescence lateral chromatogra- microspheres and antibody molecules. EDAC: Carbodiimide. –COOH:

phy; ABAD: Automatic biochemical analyzer detection. Carboxyl. –NH2: Amidogen. IgG: Immunoglobulin G.

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fluorescence-labeled binding, after which polyester fiber Table II. Comparison of hsCRP and PCT by quantitative detection film was used as a solid phase carrier for curing. Next, technique of immunofluorescence lateral chromatography in two groups the diluted CRP and PCT fluorescently labeled conjuga- (x ± s). tion was sprayed and dried on the glass fiber membrane Group hsCRP (mg/L) PCT (g/L) to prepare the marker pad [8]. (3) Antibody coating. The Infected group (n = 6) 52.32 ± 1.12 0.72 ± 0.05 monoclonal antibody CRP-C2 was selected, and the pre- Aon-infected group (n = 58) 18.75 ± 0.48 0.16 ± 0.01 adjusted coated buffer (including trehalose, tween-20 and t 139.900 76.220 buffer) was used as the coated matrix. Finally, CRP and P <0.001 <0.001 PCT capture antibodies were diluted with buffer solution, sprayed with XYZ3060 spray point apparatus (BioDot, USA), and dried on nitric acid fiber membrane to prepare group were also significantly higher than the non-infected the coated membrane [9]. group (P<0.05) (Table III, Figs. 3(C, D)).

2.2.2. Detection of hsCRP Content 3.2. Comparison of the Efficacy of the Two Detection Two to five milliliters venous blood was collected in the Methods for hsCRP and PCT Content morning from 12 h-fasting patients and centrifuged for The IFLC results showed that the AUC, sensitivity, and 5 min at 3,000 rpm to obtain serum, which was imme- specificity of hsCRP were higher than PCT. The AUC, sen- diately stored at −80 C for freezing preservation before sitivity and the jointly detected specificity of hsCRP+PCT preparation for detection within 2 h. The blood sample was were significantly higher than the individually detected mixed with diluent in a ratio of 1:10 and coated on the hsCRP and PCT (P<0.05) (Table IV, Fig. 4(A)). sample pad. After standing for 15 min, the fluorescence The BA results showed that the AUC, sensitivity, and intensity of the T line (HT) and C line (HC) was observed. specificity of hsCRP were all higher than PCT. The AUC, sensitivity, and specificity of the jointly detected

2.2.3. Detection of PCT Content Article The procedure used for the detection of the PCT content was the same as hsCRP. However,IP: 192.168.39.210 the PCT monoclonal On: Sun, 26 Sep 2021 07:58:46 antibody 16B5 was used to conjugateCopyright: the ﬂuorescent American Scientific Publishers microspheres in the preparation of CM-EUS strips,Delivered and by Ingenta PCT monoclonal antibody 42 was used as the coated antibody.

2.2.4. Automatic Biochemical Analyzer Detection The BA was used to detect hsCRP and PCT in all patients. The hsCRP content was detected by Roche Cobas 8000C701 BA, and the content of PCT was detected by Roche E411 luminescence analyzer.

2.3. Statistical Processing Method We used the SPSS 20.0 statistical software to process the data. The quantitative data were expressed as “x ± s”and analyzed by the t-test. The qualitative data were repre- sented by percentage (n) and analyzed by 2 detect. Diag- nostic performance parameters including the area under the curve (AUC), sensitivity and specificity were analyzed by logistic regression analysis and ROC curve. P<0.05 was considered statistically significant. Fig. 3. Comparison of hsCRP and PCT in two groups by quantitative detection technique of immunofluorescence lateral chromatography and automatic biochemistry analyzer. (A) Comparison of hsCRP 3. RESULTS AND DISCUSSION in two groups by quantitative detection of immunofluorescence lateral 3.1. Comparison of hsCRP and PCT in Two chromatography (P<0.05). (B) Comparison of PCT in two groups Groups by IFLC and BA by quantitative detection technique of immunofluorescence lateral chromatography (P<0.05). (C) Comparison of hsCRP in two groups When measured by IFLC, the hsCRP and PCT levels in by automatic biochemical analyzer detection, (P<0.05). (D) Compar- the infected group were significantly higher than the non- ison of PCT in two groups by automatic biochemical analyzer detec- infected group (P<0.05) (Table II, Figs. 3(A, B)). Mea- tion, (P<0.05). IG: Infected group. NG: Non-infected group. hsCRP: sured by BA, the hsCRP and PCT levels in the infected Hypersensitive C-reactive protein. PCT: Procalcitonin.

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Table III. Comparison of hsCRP and PCT detection by automatic biochemistry analyzer in two groups (x ± s).

Group hsCRP (mg/L) PCT (g/L)

Infected group (n = 6) 41.21 ± 1.05 0.61 ± 0.04 Aon-infected group (n = 58) 13.75 ± 0.48 0.15 ± 0.03 t 116.800 34.680 P <0.001 <0.001

hsCRP + PCT were significantly higher than the individually detected hsCRP and PCT (P<0.05) (Table V, Fig. 4. Comparison of the efficacy of two detection methods for Fig. 4(B)). hsCRP and PCT content. (A) Quantitative detection technique of A comprehensive analysis found that the AUC, sensi- immunofluorescence lateral chromatography; (B) automatic biochem- tivity, and the jointly detected specificity of hsCRP + PCT istry analyzer detection. The diagnostic efficiency of the two detec- by IFLC were significantly higher than obtained using the tion methods for hsCRP and PCT, the AUC, sensitivity and specificity + BA (P<0.05). of joint detection of hsCRP PCT were significantly higher than hsCRP and PCT detected individually (P<0.05). hsCRP: Hypersensi- tive C-reactive protein. PCT: Procalcitonin. 3.3. Discussion COVID-19 is a viral infection that may cause acute pneumonia. Most patients obtain a good prognosis after treat- (P<0.05). These results also indicated significant dif- ment, and only a small proportion of severe cases develop ferences in hsCRP and PCT levels between the patients septic shock, acute respiratory distress syndrome, or die. with mental illness with and without COVID-19 infec- The pathogenic virus is primarily transmitted during the tion. HsCRP is an acute phase protein synthesized by incubation period of 1–14 d through micro-droplets and liver cells. The concentration of hsCRP increased signifi- close contact, and according to the results of an epi- cantly within 2 h after the body produced an inflammatory demiological investigation [10],IP: the 192.168.39.210 general population On: is Sun,response. 26 Sep 2021 In patients 07:58:46 with mental illness and COVID-19 commonly susceptible to COVID-19.Copyright: Patients with American mental Scientificinfection, Publishers hsCRP increased sharply and then decreased Delivered by Ingenta diseases are more likely to have heightened reactions of slowly, reflecting the prognostic trend of patients [12]. fear, anxiety, and depression due to their neurological dis- Therefore, hsCRP levels in patients with COVID-19 infection increased rapidly, which is of application value for Article order, lack of prevention, and compromised psychological coping methods. Therefore, their psychological problems the early diagnosis, dynamic monitoring and prognostic caused by COVID-19 are often more serious and diffi- assessment of the novel Coronavirus. COVID-19 is a new cult to treat. Consequently, a rapid and effective test for infectious disease that can be transmitted from person to COVID-19 detection is a matter of great significance for person. It is characterized by rapid transmission and high people with mental disease. infectivity and poses a serious threat to national health. In this study, IFLC was used for detection. First, hsCRP According to epidemiological studies, COVID-19 patients, and PCT monoclonal antibodies were screened for pair- their families, and the general public exhibit adverse psy- ing, then, a fluorescence-labeled binding compound was chological problems to varying degrees [13]. The cogni- prepared with the captured antibody to form a double- tive, behavioral, and other mental facilities of mentally antibody sandwich system, and finally, hsCRP and PCT ill patients are impaired. Their ability to prevent COVID- samples from serum were combined for detection. The 19 infection is compromised, the adverse effect of the hsCRP level of normal adults is 0.068–8.2 mg/L (posi- psychological burden of COVID-19 is greater, and their tive threshold ≥5 mg/L), and the normal PCT range is ability to self-regulate is lower than non-mentally ill indi- <0.15 g/L (positive threshold >0.5 g/L) [11]. This study viduals. The application of IFLC for the rapid, precise, showed that hsCRP and PCT levels in the infected group and clinically early detection of hsCRP levels in mental were significantly higher than the non-infected group illness patients with COVID-19 will ensure that patients

Table IV. Diagnostic efﬁciency of immunoﬂuorescence lateral chromatography for determination of hsCRP and PCT.

Variables AUC 95% CI Standard error P Critical value Sensitivity (%) Speciﬁcity (%)

hsCRP 0.850 0.735–0.965 0.059 0.020 0.733 100.0 73.3 PCT 0.813 0.595–1.000 0.102 0.038 0.583 75.0 83.3 hsCRP joint PCT 1.000 1.000–1.000 0.000 0.001 1.000 100.0 100.0

Note: CI was for conﬁdence interval.

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Table V. Diagnostic efﬁciency of automatic biochemistry analyzer for determination of hsCRP and PCT.

Variables AUC 95% CI Standard error P Critical value Sensitivity (%) Speciﬁcity (%) hsCRP 0.813 0.689–0.936 0.063 0.038 0.750 100.0 75.0 PCT 0.775 0.067–0.575 0.102 0.067 0.450 100.0 45.0 hsCRP joint PCT 0.831 0.000–1.000 0.154 0.027 0.750 87.4 100.0

Note: CI was for confidence interval. are isolated as early as possible and provided with effec- fluorescence-labeled polymer had higher stability and sen- tive psychological counseling to avoid secondary mental sitivity, and the signal cascade amplification efficiency injury. Early detection can promote the rehabilitation of was stronger, which could promote the formation of the COVID-19 disease; therefore, the use of IFLC for early double-antibody sandwich system; this was combined with and accurate diagnosis of patients with mental illness and the use of coupling buffer, sample diluent, and activation COVID-19 is of great significance. PCT is a glycopro- buffer, to achieve the joint detection of hsCRP and PCT tein, a precursor substance formed by calcitonin, with no levels. Due to individual differences and limited method- hormonal activity; its content is not usually accompanied ologies, the clinical diagnosis of COVID-19 infection by by the occurrence or progression of viral infection and single detection tests is often prone to misdiagnosis, and only increases after 24 h of infection, but not significantly. missed diagnosis. The detection of the hsCRP and PCT However, in the diagnosis of COVID-19 infection, even levels by IFLC could greatly improve the accuracy and if PCT is negative, it has a certain suggestive value [14]. efficiency of diagnosis and guide the rational selection of According to the data, the PCT content increased signifi- antibiotics in clinical practice. Therefore, the combined cantly in bacterial infection. Severe sepsis, bacterial infec- detection of hsCRP and PCT by ILFC helps identify and tion, and organ function damage can lead to a significant diagnose mental diseases with COVID-19 infection and increase in serum PCT content. However, in viral infection, has a high clinical application value. Article autoimmune, and allergic diseases, the PCT content does not increase significantly [15]. Therefore,IP: 192.168.39.210 PCT content On: can Sun,Ethical 26 Sep Compliance 2021 07:58:46 be used as a reliable, acute parameterCopyright: to exclude American the pos- ScientificResearch Publishers experiments conducted in this article with ani- sibility of bacterial pneumonia in the early diagnosisDelivered of bymals Ingenta or humans were approved by the Ethical Committee pneumonia infection, which to a certain extent, is con- and responsible authorities of our research organization(s) ducive to the early clinical judgment of COVID-19. Rele- following all guidelines, regulations, legal, and ethical vant studies have shown that PCT is a landmark substance standards as required for humans or animals. for the differentiation of viral and bacterial infections [16]. In this study, the AUC, sensitivity and combined detec- Conflicts of Interest tion of the specificity of hsCRP + PCT in both the IFLC There are no conflicts to declare. and BA detection methods were better than the individual detection of hsCRP and PCT. The combined detec- Acknowledgment: Hospital level project of Shanghai tion of hsCRP and PCT by IFLC showed a diagnostic Mental Health Center (2016-YJ-09); Characteristic dis- AUC of 1.000 and a sensitivity and specificity of 100.0%. cipline construction project of Shanghai Mental Health This was superior to hsCRP, PCT single detection, and Center (2017-TSXK-07); Wu Jieping Medical Founda- hsCRP + PCT combined detection using the BA. With tion (320675015232); Chinese medicine research project IFLC, diagnostic efficiency is greatly improved and has a of Shanghai Health and Family Planning Commission in greater clinical diagnostic significance. In this study, poly- 2018 (2018LP024). mers such as PVP, PVA, tween-20 and triton X-100 were added to the detection process to improve the dispersion References and Notes and release level of the labeled binder, and the addition of 1. Byass, P., 2020. Eco-epidemiological assessment of the COVID- BSA can reduce the non-specific adsorption interference 19 epidemic in China, January-February 2020. Global Health of the labeled binder, both of which can improve the accu- Action, 13(1), p.1760490. racy of detection. Thus, the target protein can be rapidly 2. Regev, S. and Josman, N., 2020. Evaluation of executive functions detected within 15 min, with a high diagnostic value [17]. and everyday life for people with severe mental illness: A systematic review. Schizophrenia Research: Cognition, 21, p.100178. 3. Shen, L., Huang, F., Chen, X., Xiong, Z., Yang, X., Li, H., Cheng, F., 4. CONCLUSION Guo, J. and Gong, G., 2020. Journal of Zhejiang University (Medical sciences), 49(2), pp.185–190. In this study, fluorescence nano-microspheres were used 4. Qie, Z., Yan, W., Gao, Z., Meng, W., Xiao, R. and Wang, S., as tracers in the IFLC quantitative detection tech- 2019. Ovalbumin antibody-based fluorometric immunochromato- nique. Compared with the conventional luciferin, the graphic lateral flow assay using CdSe/ZnS quantum dot beads as

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