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[从中耳炎积液中分离和鉴定衣原体属]

[Isolation and identification of Chlamydia spp. from effusion of otitis media].

作者信息

Bannai H, Komoda T, Miyazawa H, Ashihara Y, Hagiwara T, Shiga S

机构信息

Department of Public Health, Kyorin University School of Health Sciences.

出版信息

Kansenshogaku Zasshi. 1991 Dec;65(12):1606-13. doi: 10.11150/kansenshogakuzasshi1970.65.1606.

DOI:10.11150/kansenshogakuzasshi1970.65.1606
PMID:1783812
Abstract

Three chlamydial strains isolated from patients of otitis media with effusion were studied by comparing reactivity to monoclonal antibody (MAb) and polyclonal antibody (PAb) produced against one clinical isolate (named Mk), which was first isolated by Dr Mukai (Mukai Microbiological Research Laboratory, Yamato-shi, Kanagawa prefecture). Commercially supplied antibody (Microtrak (Syva), Culture-set (Ortho diagnostic system)) was also used. To isolate the Chlamydia spp, the yolk sacs of eggs were immediately inoculated with sample effusions (0.2 to 0.4 ml per sac) as soon as the samples were received. The eggs were observed every day for a period of 12 days thereafter for signs of life or death. One to two blind passages were first done in the eggs and then in HeLa 229 cells. The reactivity was examined by both micro-IF tests, among various strains of Chlamydia (C. trachomatis: L2. C. pneumoniae, C. psittaci: Budgerigar, Izawa, Meningopneumonitis (MP)) and by immunoblot analysis. Chlamydia spp were isolated in two of the twenty-nine sample effusions (6.9%). These isolates were then tested for reactivity to MAb and PAb. It was found that MAb reacted with MP and Mk, but not with Budgerigar, Izawa and C. pneumoniae. The antibody of Culture-set reacted with C. trachomatis C. pneumoniae and C. psittaci. No reactivity was observed in Mk by MicroTrak. Immunoblot analysis revealed that MAb reacted with about 95 KDa protein of Mk, the two clinically isolated Chlamydia spp and MP. By using PAb from rabbits, similar blotting patterns were observed in Mk, the clinical isolates and MP.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

对从分泌性中耳炎患者中分离出的三株衣原体菌株进行了研究,通过比较它们与针对一株临床分离株(命名为Mk)产生的单克隆抗体(MAb)和多克隆抗体(PAb)的反应性,该临床分离株最初由Mukai博士(神奈川县大和市Mukai微生物研究实验室)分离得到。还使用了市售抗体(Microtrak(Syva公司)、Culture-set(Ortho诊断系统公司))。为了分离衣原体属,样本积液一收到就立即接种到鸡蛋的卵黄囊中(每个卵黄囊接种0.2至0.4毫升)。此后每天观察鸡蛋12天,观察其存活或死亡迹象。首先在鸡蛋中进行一到两次盲传,然后在HeLa 229细胞中进行盲传。通过微量免疫荧光试验在各种衣原体菌株(沙眼衣原体:L2、肺炎衣原体、鹦鹉热衣原体:虎皮鹦鹉株、Izawa株、脑膜肺炎株(MP))中检测反应性,并通过免疫印迹分析进行检测。在29份样本积液中有两份分离出了衣原体属(6.9%)。然后对这些分离株进行MAb和PAb反应性检测。发现MAb与MP和Mk反应,但不与虎皮鹦鹉株、Izawa株和肺炎衣原体反应。Culture-set抗体与沙眼衣原体、肺炎衣原体和鹦鹉热衣原体反应。MicroTrak在Mk中未观察到反应性。免疫印迹分析显示,MAb与Mk、两株临床分离的衣原体属菌株和MP的约95 kDa蛋白反应。使用兔源PAb时,在Mk、临床分离株和MP中观察到类似的印迹模式。(摘要截短于250字)

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