Preparation of soluble microneedle patch with fusion protein nanoparticles secreted by <italic>Mycobacterium tuberculosis</italic> and application of tuberculosis skin test

Fan CHEN; Rong-sheng ZHU; Jing ZHOU; Yue HU; Yun XUE; Jian-hua KANG; Wei WANG

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Preparation of soluble microneedle patch with fusion protein nanoparticles secreted by Mycobacterium tuberculosis and application of tuberculosis skin test

VernacularTitle:结核分枝杆菌分泌蛋白融合蛋白纳米粒可溶性微针贴片的制备及结核病皮试的应用
Author: Fan CHEN ¹ ; Rong-sheng ZHU ¹ ; Jing ZHOU ² ; Yue HU ¹ ; Yun XUE ³ ; Jian-hua KANG ⁴ ; Wei WANG ⁴
Author Information

1. State Key Laboratory of Biocatalysis and Enzyme Engineering, Life Science School of Hubei University, Wuhan 430062, China
2. Clinical Laboratory of Wuhan Pulmonary Hospital, Wuhan 430030, China
3. College of Medical Technology and Engineering, Henan University of Science and Technology, Luoyang 471003, China
4. Department of Laboratory Medicine, Dongguan Chang′an Hospital, Dongguan 523843, China
Publication Type:Research Article
Keywords: tuberculosis; skin test; nanoparticle; microneedle patch
From: Acta Pharmaceutica Sinica 2024;59(6):1804-1811
CountryChina
Language:Chinese
Abstract: Rapid epidemiological screening for tuberculosis (TB) usually uses tuberculin pure protein derivative (PPD) skin test, which has limitations such as low specificity and high side effects. ESAT-6 and CFP-10 are secreted proteins of Mycobacterium tuberculosis, but the related genes are missing from Bacillus Calmette-Guerin (BCG). In this study, the fusion protein ESAT6-CFP10 (EC) was expressed and purified, and prepared into chitosan nanoparticles (EC-NPs), which were loaded into the microneedle patch and carried out the preliminary test of tuberculosis skin test. The drug loading capacity of MNP-EC-NPs (microneedle patch, MNP) can reach 0.03 μg per needle and 1.92 μg per patch. The shelf life of MNP-EC-NPs can reach 6 months at room temperature, and it can effectively penetrate the epidermis. Volunteer skin test results showed that MNP-EC-NPs can effectively distinguish between BCG vaccinators, and can effectively show positive reaction in the skin of tuberculosis patients without significant side effects. The experiment was approved by the Ethics Committee of Wuhan Pulmonary Hospital [2022 (2)]. In this study, a TB skin test method was established, using ESAT6-CFP10 fusion protein instead of PPD, and using soluble microneedle dosage form, which improved the specificity of TB skin test diagnosis and provided a new technical scheme for TB epidemic screening.