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Accurate Detection of Streptococcus pneumoniae by Using Ribosomal Protein L7 / L12 as Molecular Marker |
WANG Meng1,SONG Hui-ru1,CHENG Yu-jie1,WANG Yi1,2,3,YANG Bo1,2,3,HU Zheng1,2,3,**() |
1 School of Food and Biological Engineering, Hubei University of Technology, Wuhan 430068, China 2 Key Laboratory of Fermentation Engineering (Hubei University of Technology), Ministry of Education, Wuhan 430068, China 3 Hubei Collaborative Innovation Center for Industrial Fermentation, Wuhan 430068, China |
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Abstract Objective: A rapid colloidal gold immunochromatography assay (GICA) for the detection of Streptococcus pneumoniae was explored by using the ribosomal protein L7/L12 as detection marker. Method: The gene sequence of ribosomal protein L7/L12 was analyzed, the prokaryotic expression vector was constructed, the recombinant protein was expressed and purified, and the monoclonal antibody was prepared by immunizing BALB/c mice; The affinity of antigens and antibodies was detected by a labelless molecular interaction instrument based on BLI technology; The colloidal gold immunochromatographic test strip was developed based on the principle of double antibody sandwich, and its specificity, sensitivity and stability were evaluated. Result: Two hybridoma cell strains that can efficiently secrete anti-monoclonal antibody were acquired by means of screening. Purified monoclonal antibodies all had high affinity and no competition with antigen. Pairing and detection were made on colloidal gold immunochromatography platform. The minimum detection limit of the test strips was 1.0×105 CFU/ml; It had a specific reaction with Streptococcus pneumoniae, but did not cross-react with other 9 kinds of common respiratory pathogens such as Haemophilus influenzae and Moraxella catarrhalis; Test strips were kept at 25°C for 12 months and still had good repeatability and stability. Conclusions: RP-L7/L12 can be used as a detection marker for Streptococcus pneumonia. The colloidal gold immunochromatographic test strips prepared by its monoclonal antibody can be used for rapid detection of streptococcus pneumoniae.
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Received: 10 October 2019
Published: 18 May 2020
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Corresponding Authors:
Zheng HU
E-mail: zhenghu@mail.hbut.edu.cn
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