水溶性对硝基苯甲酸在结核分枝杆菌菌种鉴定中的应用

Application of Water-soluble P-nitrobenzoic Acid at the Mycobacterium Identification of Bacteria

目的探讨分枝杆菌(Mycobacterium)菌群鉴定中应用水溶性对硝基苯甲酸(P-Nitro benzoic Acid,PNB)的可行性。方法用0.1mol/LNaoH来溶解对硝基苯甲酸基(即水溶性PNB),替代用传统的方法(有机溶剂溶解法)溶解对硝基苯甲酸。在改良罗氏培养基中,对22种标准菌株水溶性对硝基苯甲酸用三种浓度:0.25mg/ml,0.5mg/ml,1.0mg/ml,有机溶剂用规程要求的浓度:0.5mg/ml进行菌型鉴定的对照;100株临床分离菌株分别用两种溶解方法制备的含0.5mg/mlPNB的培养基进行菌型鉴定的对照。结果 22种标准菌株,在终浓度为0.5mg/ml时,水溶性PNB和用二甲亚枫溶解的PNB在对分枝杆菌的鉴定结果相同。100株临床分离菌株两种方法的结果均为:结核分枝杆菌82例,非结核分枝杆菌18例,结果完全相同。结论用0.5mg/ml的水溶性对硝基苯甲酸是完全可以替代传统的有机溶剂溶解方法的,并且在液体培养基中可避免有机溶剂溶解的PNB引起蛋白沉淀而影响试验结果。

Objective To discuss the application feasibility of water-soluble p-nitrobenzoic acid at the Mycobacterium Identification of bacteria. Methods 0.1 mol/L NaoH to dissolve p-nitrobenzoic acid （water-soluble PNB） was used to replace the traditional methods （organic solvent dissolution method） to dissolve the p-nitrobenzoic acid. In Lowenstein-Jensen culture medium, 22 standard strains used three concentrations of water-soluble p-nitrobenzoic acid： 0.25 mg/ml, 0.5 mg/ml, 1.0 mg/ml, organic solvent concentration required to use a point of order： 0.5 mg/ml as control at identification of bacteria; 100 clinical isolates were prepared using two different dissolution medium containing 0.5mg/ml PNB. Results The identification of 22 standard strains was the same, using water-soluble PNB and PNB dissolved with Dimethyl sulfoxide at the final density 0.5 mg/ml. 100 clinical separation strains used two methods： The mycobacterium tuberculosis 82 cases, the non-mycobacterium tuberculosis 18 cases, the result was completely same. Conclusions The 0.5 mg/ml water-soluble p-nitrobenzoic acid may replace the traditional organic solvent dissolution method entirely, and may avoid PNB which in the liquid media the organic solvent dissolves causing the protein to precipitate affects the test result.