Differentiation of Bartonella-like isolates at the species level by PCR-restriction fragment length polymorphism in the citrate synthase gene

• Published in: Journal of Clinical Microbiology 1995-07, Vol.33 (7), p.1797-1803
• Main Authors: Norman, A.F. (University of Georgia, Athens, GA.), Regnery, R, Jameson, P, Greene, C, Krause, D.C
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Bacteriological methods and techniques used in bacteriology; Bacteriology; Bartonella - enzymology; Bartonella - genetics; Bartonella - isolation & purification; Bartonella henselae; Bartonella henselae - enzymology; Bartonella henselae - genetics; Bartonella henselae - isolation & purification; Bartonella quintana; Base Sequence; Biological and medical sciences; CAROLINA DEL NORTE; CAROLINE DU NORD; Cat-Scratch Disease - diagnosis; Cat-Scratch Disease - microbiology; Cats; CHAT; Citrate (si)-Synthase - genetics; CLONACION; CLONAGE; Cloning, Molecular; DNA Primers - genetics; DNA, Bacterial - genetics; Fundamental and applied biological sciences. Psychology; GATO; GENE; GENES; Genes, Bacterial; Genotype; Humans; Indicators and Reagents; Microbiology; Molecular Sequence Data; PCR; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - statistics & numerical data; Polymorphism, Restriction Fragment Length; Restriction Mapping; RFLP; RICKETTSIALES; SECUENCIA NUCLEOTIDICA; Sequence Homology, Nucleic Acid; SEQUENCE NUCLEOTIDIQUE; Species Specificity; VARIACION GENETICA; VARIATION GENETIQUE
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/jcm.33.7.1797-1803.1995

The citrate synthase gene (gltA) of Bartonella henselae was cloned and sequenced to compare genetic divergence among alpha and gamma branches of the class Proteobacteria and to develop enhanced genotypic reagents for B. henselae identification. B. henselae gltA is 1,293 nucleotides in length and 63 to 66% homologous with corresponding gene sequences of Rickettsia prowazekii, Escherichia coli, and Coxiella burnetii. The observed genetic variability suggests that gltA sequences can provide a useful means for studying moderate divergence among related bacteria. Oligonucleotides specific for B. henselae gltA were evaluated for the ability to prime PCR amplification within the alpha and gamma branches of the proteobacteria. Under the conditions used, only B. henselae, Bartonella quintana, and R. prowazekii template DNAs yielded amplification products (approximately 380 bp). DNAs from 28 Bartonella-like isolates of feline origin were amplified by B. henselae primers and analyzed for restriction fragment length polymorphism. The resulting patterns for all 28 isolates were similar or identical to that of the recognized B. henselae strain. Current studies are aimed at optimization of PCR conditions for specificity and sensitivity of amplification of Bartonella sequences from clinical isolates