Reverse line blot assay for direct identification of seven Streptococcus agalactiae major surface protein antigen genes.
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 (Calpha), Rib, Epsilon (Epsilon/Alp1/Alp5), Alp2, Alp3, and Alp4, and the immunoglobulin A binding protein, Bac (Cbeta). 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.
Zhao
Zuotao
Z
Centre for Infectious Diseases and Microbiology (CIDM), Institute of Clinical Pathology and Medical Research (ICPMR), Westmead Hospital, Darcy Road, Westmead, New South Wales 2145, Australia.
Kong
Fanrong
F
Gilbert
Gwendolyn L
GL
D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D000954:Antigens, Surface / Q000235:genetics* / Q000378:metabolism; D017403:In Situ Hybridization / Q000379:methods; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012703:Serotyping; D013292:Streptococcus agalactiae / Q000235:genetics* / Q000276:immunology*
D000942:Antigens, Bacterial; D000954:Antigens, Surface