Typing of Haemophilus parasuis strains by PCR–RFLP analysis of the tbpA gene

• Published in: Veterinary microbiology Vol. 92; no. 3; pp. 253 - 262
• Main Authors: de la Puente Redondo, V.A, Navas Méndez, J, Garcı́a del Blanco, N, Ladrón Boronat, N, Gutiérrez Martı́n, C.B, Rodrı́guez Ferri, E.F
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 02-04-2003; Elsevier Science
• Subjects: Animals; Bacteriology; Biological and medical sciences; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; Fundamental and applied biological sciences. Psychology; Genetics; Haemophilus - chemistry; Haemophilus - classification; Haemophilus - genetics; Haemophilus Infections - microbiology; Haemophilus Infections - veterinary; Haemophilus parasuis; Microbiology; PCR–RFLP; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - veterinary; Polymorphism, Restriction Fragment Length; Sensitivity and Specificity; Swine; Swine Diseases - microbiology; tpbA gene; Transferrin-Binding Protein A - chemistry; Transferrin-Binding Protein A - genetics; tpbA gene; PCR–RFLP; Haemophilus parasuis; Pasteurellaceae; Polymerase chain reaction; Restriction fragment length polymorphism; Typing; Haemophilus; Bacteria; PCR-RFLP
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/S0378-1135(02)00362-0

On the basis of a species-specific PCR assay, a RFLP analysis for typing of
Haemophilus parasuis strains was developed and evaluated. Amplification was based on the gene
tbpA, encoding a transferrin-binding protein. RFLP analysis of the 1.9-kb
tbpA-amplicon using
TaqI,
AvaI and
RsaI endonucleases produced 12 different patterns for the reference strains of the 15 known
H. parasuis serovars, and showed a high heterogeneity (33 RFLP groups) for 101
H. parasuis clinical isolates tested. The sensitivity, typeability (100% versus 65% for immunodiffusion), high degree of discrimination (0.93 versus 0.84 for immunodiffusion), simplicity and low cost per test make this PCR–RFLP assay a useful method for typing
H. parasuis and, therefore, for studying the epidemiology of outbreaks of Glässer’s disease.