Primer for detecting serotype of shigella flexneri and multiplex amplification using same

Abstract

The invention relates to a primer for detecting the serotype of shigella flexneri, and the method comprises the following sequences: SEQ ID Nos. 1 and 2 (sequence identity numbers 1 and 2), SEQ ID Nos. 3 and 4, SEQ ID Nos. 5 and 6, SEQ ID Nos. 7 and 8, SEQ ID Nos. 9 and 10, SEQ ID Nos. 11 and 12, SEQ ID Nos. 13 and 14, and SEQ ID Nos. 15 and 16. The primer has specificity and consistent annealingtemperature. The invention further relates to a method for performing multiplex amplification by utilizing the primer. The invention further relates to an application of the primer in detecting the serotype of the shigella flexneri in the preparation of detection agents. The invention further relates to a kit containing the primer for detecting the serotype of the shigella flexneri.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to primers for Shigella flexneri detection and multiple amplification using the primers. Background technique [0002] Shigella is the main pathogen of bacterial diarrhea in developing countries. Each year, 164.7 million people worldwide are infected, resulting in 110,000 deaths, mostly children under the age of 5 (Kotloff, K.L., J.P. Winickoff, B. Ivanoff, J.D. Clemens, D.L. Swerdlow, P.J. Sansonetti, G.K. Adak, and M.M. Levine. 1999. Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull World Health Organ 77:651-66). Of the four groups of Shigella, Flexneri is the predominant group affecting the poor. [0003] According to the structure of O-antibody, Shigella flexneri is divided into different serotypes. So far, at least 15 serotypes have been reported, namely 1a, 1b, 1c, 2a, 2b, 3a, 3b, 4a, 4b, 5a, 5b, X, Xv, Y, ...

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Problems solved by technology

However, this method has some disadvantages. First, the slide agglutination method to determine the serotype of a strain of Flexneri requires up to 10 reaction tests, respectively with type antigens I, II, III, IV, V, VI, group antigen 3 ; 4, 7; 8, 6 and the monoclonal antibody MASFlc against serotype 1c (Stagg, R.M., P.D. Cam, and N.K. Verma. 2008. Identification of newly recognized serotype 1c as the most prevalent Shigella flexneri serotype in northern rural Vietnam. Epidemiol Infect 136: 1134-40; Talukder, K.A., Z. Islam, M.A. Islam, D.K. Dutta, A. Safa, M. Ansaruzzaman, A.S. Faruque, S.N. Shahed, G.B. Nair, and D.A. Sack. 2003. Phenotypic and genotypic characterization of provisional serotype Shigella flexneri 1c and clonal relationships with 1a and 1b strains isolated in Bangladesh. J Clin Microbiol 41:110-7; Ye, C., R. Lan, S. Xia, J. Zhang, Q. Sun, S. Zhang, H. Jing, L. Wang, Z. Li, Z. Zhou, A. Zhao, Z. Cui, J. Cao, D. Jin, L. Huang, Y. Wang, X. Luo, X. Bai, P. Wang, Q.Xu, and J.Xu.2010.Emergence of a new multidrug-resistant serotype X variant in an epidemic clone of Shigella flexneri.J Clin Microbiol 48:419-26); Interpretation of results can lead to false results; finally, expensive antiserum kits limit the use of this method in laboratories in developing countries

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