Attempt to develop a simple and rapid method for diagnosis of cat scratch disease (Bartonella infection).

1.对汉氏巴尔通体全菌进行了SDS-PAGE分析，发现有35条以上的蛋白质条带。主要蛋白质的相对分子质量分别为90、62、45(43)、40、35、31和25 kDa。主要外膜蛋白的相对分子质量分别为62、43和31kDa。108例有淋巴结肿大怀疑为慢性阻塞性肺病的患者中，间接免疫荧光试验(IFA)检测到32例抗体滴度超过256倍。用病人血清对氏杆菌全细胞进行免疫印迹分析，发现有8条带(90、65、43、40、35、33、30和28 kDa)。阳性条带的分布模式因病例而异，但65、43和35 kDa蛋白具有较强的抗原性。与Vero细胞共培养可提高IFA的反应性。这种反应性的增加在35 kDa条带上表现得非常明显，但有些血清不与该条带反应。因此，我们找不到可用于血清学诊断的单一抗原蛋白。包括65、43和35 kDa蛋白在内的复合蛋白可作为CSD实验室诊断的新方法。4.B.henselae可刺激淋巴管内皮细胞和血管内皮细胞的生长。亨氏假单胞菌可诱导中性粒细胞表面CCR7表达。这种细菌还能刺激淋巴管内皮细胞产生CCL21。然后我们检测了CCL21对慢性阻塞性肺疾病患者的诊断价值。结果表明，CSD患者血清CCL21水平与其他不同淋巴结病患者无显著差异。5.我们建立了一种新的亨塞氏杆菌培养方法，并用该方法从流浪猫体内分离到16株亨塞氏杆菌。

1.Analysis by SDS-PAGE of whole bacterial cells of Bartonella henselae revealed more than 35 protein bands. The molecular weight of major proteins were 90, 62, 45 (43), 40, 35, 31, and 25kDa. Molecular weight of the major outer membrane proteins were estimated as 62, 43 and 31kDa.2.Thirty-two of 108 patients who had swelling of lymph nodes and CSD was suspected showed more than 256x IgG titer against B.henselae by indirect immunofluorescence assay (IFA). These antibody positive sera were used for the following study.3.Western blotting analysis of whole bacterial cells of B henselae using patient sera revealed 8 bands (90, 65, 43, 40, 35, 33, 30, and 28kDa). Distribution pattern of positive bands were diverse in case to case, however, 65, 43, and 35kDa proteins showed strong antigenicity. The reactivity of IFA increased by co-cultivation of B.henselae with vero cells. This increase of reactivity were striking in 35kDa band, however, some sera did not reacted with this band. Consequently we could not find sole antigen protein useful for sero-diagnosis. Compound proteins, including 65, 43, and 35kDa protein, should be considered to develop new method for the laboratory diagnosis of CSD.4.B.henselae stimulated the growth of lymphatic endothelial cells as well as vascular endothelial cells. B.henselae induced CCR7 on the surface of peripheral neutrophils. The bacteria also stimulated the production of CCL21 from lymphatic endothelial cells. Then we examined the diagnostic importance of CCL21 in patients with CSD. The result showed no significant difference of serum CCL21 level between CSD and other patients with various lymph node diseases.5.We developed a new method to cultivate B.henselae, and isolated 16 strains of B.henselae from stray cats by using the method.