Development of a Colloidal Gold Immunochromatographic Test Strip for the Detection of Moraxella catarrhalis

doi:10.13560/j.cnki.biotech.bull.1985.2018-0184

Abstract

Abstract: The objective of this work is to develop a colloidal gold immunochromatographic test strip for the expeditious detection of Moraxella catarrhalis in the respiratory tracts of infected patients,and then to establish the corresponding approach. By bioinformatics analysis,the linear antigenic epitope on UspA1 of Moraxella catarrhalis was discovered and named UspA1Line1. The UspA1Line1 protein then ligated with KLH to generate the UspA1Line1-KLH protein. The already-constructed engineering strain for expressing UspA1 was used to express and purify the recombinant UspA1-His protein,which then was used to immune New Zealand rabbits for preparing the polyclonal antibody. Then the serum from the rabbits was purified by two steps of Protein A and polypeptide affinity chromatography,and the polyclonal antibody with high-purity and strong-specificity was acquired. The 40 nm colloidal gold particles were prepared by trisodium citrate reduction method,which then bound with the polyclonal antibody to form a double antibody sandwich,thus the goal of rapidly detecting M. catarrhalis was achieved. As a result,by the established approach M. catarrhalis was specifically detected in 10 minutes,and no cross-reactivity with common pathogens in upper respiratory tract occurred. The strips may be stored at 24℃ and their repeatability and stability remain well. In sum,a colloidal gold immunochromatographic test strip to rapidly detect M. catarrhalis is successfully developed.

Key words: Moraxella catarrhalis, linear antigenic epitope, UspA1, immunochromatography

HUANG Ying-qi, ZHANG Qiu, TAO Ye, HU Zheng, ZHANG Gai-ping, ZHANG Hua-shan. Development of a Colloidal Gold Immunochromatographic Test Strip for the Detection of Moraxella catarrhalis[J]. Biotechnology Bulletin, 2018, 34(9): 184-189.

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