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Clinical and Vaccine Immunology, January 2006, p. 145-149, Vol. 13, No. 1
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.1.145-149.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Reverse Line Blot Assay for Direct Identification of Seven Streptococcus agalactiae Major Surface Protein Antigen Genes

Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead, NSW 2145, Australia,¹ Department of Dermatology, First Hospital, Peking University, Beijing 100034, People's Republic of China²

Received 24 July 2005/ Returned for modification 3 October 2005/ Accepted 27 October 2005

We developed a multiplex PCR-based reverse line blot hybridization assay (mPCR/RLB) to detect the genes encoding members of the family of variable surface-localized proteins of Streptococcus agalactiae (group B streptococcus [GBS]), namely, Bca (C), Rib, Epsilon (Epsilon/Alp1/Alp5), Alp2, Alp3, and Alp4, and the immunoglobulin A binding protein, Bac (Cß). We used the assay to identify these genes in a collection of well-characterized GBS isolates and reference strains. The results showed that mPCR/RLB avoids the common problems of cross-reaction and nontypability associated with protein typing using antisera. It is as sensitive as, but more practical than, separate gene-specific PCRs and would be suitable for large molecular epidemiological studies of GBS.

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