Quantification of antigen of <i>Mycoplasma capricolum</i> subsp. <i>capripneumoniae</i> by optical assay.

Jiazhen GE; Pengcheng Gao; Tongtong Tian; Xiaoni WU; Qianqian LI; Kexin Tian; Guodong Song; Fuying Zheng; Yuefeng Chu

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Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay.

Author: Jiazhen GE ¹ ; Pengcheng GAO ¹ ; Tongtong TIAN ¹ ; Xiaoni WU ¹ ; Qianqian LI ¹ ; Kexin TIAN ¹ ; Guodong SONG ¹ ; Fuying ZHENG ¹ ; Yuefeng CHU ¹
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1. State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, Gansu, China.
Publication Type:Journal Article
Keywords: CTAB precipitation method; Mycoplasma capricolum subsp. capripneumoniae; capsular polysaccharide; modified culture-medium; ultraviolet spectrum
MeSH: Humans; Animals; Goats; Cetrimonium; Mycoplasma; Polysaccharides
From: Chinese Journal of Biotechnology 2023;39(12):4874-4886
CountryChina
Language:Chinese
Abstract: Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.