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首页> 美国政府科技报告 >Culture, Polymerase Chain Reaction and Restriction Fragment Length Polymorphism Studies on Bartonella bacilliformis
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Culture, Polymerase Chain Reaction and Restriction Fragment Length Polymorphism Studies on Bartonella bacilliformis

机译：巴氏杆菌的培养，聚合酶链反应和限制性片段长度多态性研究

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Bartonella ssp. have gained importance as etiologic agents of human disease, both in temperate and tropical regions. Reports of increasing number of clinical cases of bartonellosis in Peru (B. bacilliformis), documentation of chromic bacteremia in domestic cats with cat scratch fever(b. henselae), and the association of bacillary angiomatosis and parenchymal peliosis (B. henselace and B. quintana) in AIDs patients demand improved laboratory diagnostic detection and isolation techniques for this fastidious organism. We report successful culture and polymerase chain reaction (PCR) techniques applicable for this purpose. Lyophilized B. /bacillioformis was suspended in PUB and cultured on blood and chocolate agar plates to verify survival. Characteristic colonies were used to seed an 8% and 0.7% NaHCO3. Aliquots incubated at 28 C in a candle jar for 3-7 days showed numerous, pleomorphic intraerythrocytic bacteria when thin smears were stained with Giemsa. Ethidium bromide staining and visualization using ultraviolet light of fixed smears of washed red cells showed numerous fluorescent organisms within the cells. B. Henselae was similarly cultivated and detected after incubation at 37 C. This culture system allows for early presumptive detection of Bartonella ssp., taking advantage of the organism's predeliction for intraerythrocytic habitation and the ability to stain fixed RBCs. For PCR, primer were designed to amplify regions between the 16S and 23S rRNA genes of Bartonella, or the entire spacer region. In each case, the primers represented sequences conserved among Bartonella species, and the procedures amplified variable regions 40-1700 bp in length that should be useful for distinguishing species of Bartonella and for molecular epidemiology in restriction length polymorphism analysis.

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