马尔尼菲青霉菌素的制备和应用

目的探讨马尔尼菲青霉菌素的制备方法,并评价以马尔尼菲青霉菌素皮肤试验来诊断播散性马尔尼菲青霉菌病的价值。方法将新西兰大白兔随机分为对照组、马尔尼菲青霉菌组和白色念珠菌组,每组20只。马尔尼菲青霉菌组感染马尔尼菲青霉菌,白色念珠菌组感染白色念珠菌,对照组不作任何处理。按真菌抗原提取法制备马尔尼菲青霉菌素并进行部分改良,用Lowry法测定马尔尼菲青霉菌素中纯蛋白衍生物的含量;将马尔尼菲青霉菌素皮内注射于3组新西兰大白兔耳部内侧皮肤,并于注射后24、48、60h观察皮肤红肿反应。结果获得的马尔尼菲青霉菌素原液纯度高,特异性好;抗原效价测定结果显示马尔尼菲青霉菌素中纯蛋白衍生物含量为68mg·L-1,且具有良好的安全性。马尔尼菲青霉菌组和白色念珠菌组各时间点皮肤红肿反应平均直径比较,差别有统计学意义(P<0.001)。对照组未出现皮肤反应。注射后24h,马尔尼菲青霉菌组皮肤红肿反应全部为阳性,白色念珠菌组有2只为阳性。马尔尼菲青霉菌素皮肤试验方法对马尔尼菲青霉菌病进行早期诊断的敏感性为100.0%,特异性为90.0%,阳性预测值为90.9%,阴性预测值为100.0%。结论对真菌抗原提取法进行部分改良可获得具有良好安全性的马尔尼菲青霉菌素;马尔尼菲青霉菌素皮肤试验方法简便,结果稳定,可用于马尔尼菲青霉菌病的辅助诊断。

Preparation and application of Penicillium marneffei

Objective To discuss the preparation of Penicillium marneffei and to evaluate its clinical value in the diagnosis of Penicilliposis marneffei disease by skin test. Methods New Zealand rabbits were randomly divided into control group,Penicillium marneffi group and Candida albicans group,twenty rabbits in each group. Penicillium marneffi was infected in Penicillium marneffi group,Candida albicans was infected in Candida albicans group,no treatment was given in control group. Penicillium marneffei were prepared by partial improvement of fungal antigen extraction method,the content of purified protein derivative of Penicillium marneffei was determined by Lowry method. The inside ear skin of New Zealand rabbits in three groups were injected by endermic injection of Penicillium marneffei and the skin irritation response were observed after injection 24,48,60 hours. Results Acquired stock solution of Penicillium marneffei have high purity and good specificity. The content of purified protein derivative of Penicillium marneffei was 68 mg·L -1 and has a good safety. There was statistical significance in the average diameter of skin irritation response at each time between Penicillium marneffei group and Candida albicans group( P < 0. 001) . There was no skin irritation response happened in control group. After injection 24 hours,the skin irritation responses of all rabbits in Penicillium marneffei group were positive,and two rabbits in Candida albicans group were positive. The sensitivity,specificity,positive predictive rate and negative predictive rate of Penicillium marneffei skin test for early diagnosis of penicilliposis marneffei disease were 100. 0% ,90. 0% ,90. 9% and 100. 0% respectively. Conclusion The Penicillium marneffei which has good safety can procured by partial improvement of fungal antigen extraction method; Penicillium marneffei skin test method is simple and stable, and it may be used to the aid diagnosis of penicilliposis marneffei disease.