ESTABLISHMENT OF TIME-RESOLVED FLUORESCENCE IMMUNOCHROMATOGRAPHIC METHOD FOR TOXOPLASMA GONDII DETECTION

UDC 616.993.192.1:543.544

 

Li Laiqing – bachelor, engineer, chairman. Guangzhou Youdi Bio-technology Co., Ltd. (3, Juquan Road, Huangpu District, 510663, Guangzhou, People’s Republic of China). E-mail: lilaiqing191@163.com

Liang Huankun – bachelor, assistant engineer, quality manager. Guangzhou Youdi Bio-technology Co., Ltd. (3, Juquan Road, Huangpu District, 510663, Guangzhou, People’s Republic of China). E-mail: 605712026@qq.com

Zhong Shuhai – bachelor, assistant engineer, R&D supervisor. Guangzhou Youdi Bio-technology Co., Ltd. (3, Juquan Road, Huangpu District, 510663, Guangzhou, People’s Republic of China). E-mail: 1627840617@qq.com

Chen Cuicui – master, engineer, project manager. Guangzhou Youdi Bio-technology Co., Ltd. (3, Juquan Road, Huangpu District, 510663, Guangzhou, People’s Republic of China). E-mail: 285020003@qq.com

Leont’yev Viktor Nikolayevich – PhD (Chemistry), Associate Professor, Head of the Department of Biotechnology. Belarusian State Technological University (13a, Sverdlova str., 220006, Minsk, Republic of Belarus). E-mail: leontiev@belstu.by Voitau Ihar Vital’evich – DSc (Engineering), Professor, Rector. Belarusian State Technological University (13a, Sverdlova str., 220006, Minsk, Republic of Belarus). E-mail: voitov100@mail.ru

 

DOI: https://doi.org/ 10.52065/2520-2669-2024-277-14.

 

Key words: Toxoplasma gondii, time-resolved fluorescence microsphere, immunochromatographic assay, parasite, zoonosis.

 

For citation: Li L., Liang H., Zhong S., Chen C., Leont’yev V. N., Voitau I. V. Establishment of time-resolved fluorescence immunochromatographic method for Toxoplasma gondii detection. Proceedings of BSTU, issue 2, Chemical Engineering, Biotechnologies, Geoecology, 2024, no. 1 (277), pp. 103–110. DOI: 10.52065/2520-2669-2024-277-14.

Abstract

Toxoplasma gondii (T. gondii) is a parasitic zoonosis that causes abortion or congenital diseases. The purpose of this study is to establish a immunochromatographic assay (ICA) method using time-resolved fluorescence microspheres (TRFM) for T. gondii quantitative detection in serum. Firstly, we activated the TRFM and then coupled with anti-T. gondii antibodies (mAb1) and antiBSA-DNP antibodies to prepare the TRFM-mAb1 probes and TRFM-BSA-DNP probes. Then, we optimized the coupling pH, T line antibody concentration, probes usage and detection time. Finally, we established the T. gondii-TRFMICA method and assemble the T. gondii-TRFMICA kit, the standard curve, sensitivity, precision, specificity, clinical sensitivity and specificity were evaluated. Under the optimized conditions, the T. gondii-TRFMICA test was achieved within 20 min with the sensitivity 0.1 ng/mL. The recoveries were ranging from 100–110% with the intra-assay and inter-assay CV lower than 10%. The kits have detection specificity for positive serum of several common infectious diseases and some common serum components, and have high detection sensitivity and specificity in human and cat clinical samples. A TRFMICA kit for T. gondii quantitative detection was successfully prepared with high sensitivity, specificity, precision and clinical sensitivity and specificity. We recommend TRFMICA as a promising technique in the clinical diagnosis and monitoring of toxoplasmosis in human and animals.

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09.11.2023