制内毒素的凝集反应[6],我们用多粘菌素来抑制BC因子的活性,由于多粘菌素纯度不高,混有内毒素,结果不理想。革兰阳性菌不影响检测,革兰阴性菌影响检测。 真菌的细胞壁较为坚固,我们对真菌标准品和临床标本处理时,采用超声的方法,以破坏真菌的结构。有学者从真菌中提取出葡聚糖[7]。 G因子法已用于测定间质性浆细胞肺炎血中的葡聚糖[8],它可检测小鼠念珠菌病[9]和检测临床病人深层真菌感染[10]。 文献报道,1ng~1μg葡聚糖与G因子可产生凝集反应[11],与我们的结果相似。 我们的实验显示产生反应的葡聚糖的浓度较高,为200ng~100μg,可能与鲎试剂的敏感性不同有关。真菌或含有真菌的标本以10倍稀释,至少有3~4个浓度凝集反应显著,而细菌在8个稀释度中都有反应,另外真菌和革兰阴性细菌的时间反应曲线和时间速率曲线不同,用此方法可检测标本的真菌感染。我们将进一步分离G和B、C因子,进行特异性反应,可排除细菌对真菌检测的影响。【参考文献】 1 Thomas. Current perspectives on ophthalmic mycoses. Clin Microbiol Rev, 2003, 16:730-797. 2 王丽娅.我国真菌性角膜炎的研究现状.眼科,2005,14:143-144. 3 Salierno, Goldstein Driebe. Fungal ring infiltrates in disposable contact lens wearers. Clao J, 2001, 27:166-168. 4 Karp, Tuli, Yoo, et al. Infectious keratitis after LASIK. phthalmology, 2003, 110:503-510. 5 国家药典委员会.中华人民共和国药典,二部,附录XI E,细菌内毒素检查法,2005版,附录87. 6 Tsuchiya, Oishi, Takaoka, et al. Discrimination between endotoxin and (1~3)-beta-D-glucan using turbidimetric kinetic assay with Limulus amebocyte lysate. Chem Pharm Bull (Tokyo), 1990,38:2523-2526. 7 Williams, Mcomamee, Jones, et al. A method for the solubilization of a (1~3)-beta-D-glucan isolated from Saccharomyces cerevisiae. Carbohydr Res, 1991,219:203-213. 8 Yasuoka, Tachikawa, Shimada, et al. (1~>3) beta-D-glucan as a quantitative serological marker for Pneumocystis carinii pneumonia. Clin Diagn Lab Immunol, 1996, 3:197-199. 9 Nichterlein, Buchheidt, Hein, et al. Comparison of glucan detection and galactomannan enzyme immunoassay in gastrointestinal and systemic murine candidiasis. Diagn Microbiol Infect Dis, 2003, 46:103-108. 10 Ostrosky-Zeichner, Alexander, Kett, et al. Multicenter clinical evaluation of the (1~>3) beta-D-glucan assay as an aid to diagnosis of fungal infections in humans. Clin Infect Dis, 2005, 41:654-659. 11 Tsuchiya, Takaoka, Tokioka, et al.Development of an endotoxin-specific Limulus amebocyte lysate test blocking beta-glucan-mediated pathway by carboxymethylated curdlan and its application.Nippon Saikingaku Zasshi, 1990, 45:903-911. 上一页 [1] [2]
Tags:
|