摘要
【目的】探求一种能够快速、准确地筛选出赭曲霉毒素产生菌的分析方法,为赭曲霉毒素的检测与防治奠定基础。【方法】分别采用紫外荧光法、胶体金免疫试纸条法和高效液相色谱(HPLC)法,对分离自葡萄表面的189株霉菌产生赭曲霉毒素A(OTA)的能力进行分析,并对3种方法的鉴定结果进行比较。其中在紫外荧光法中,黑曲霉接种至可可浆培养基(CCA)培养,其他霉菌接种至察氏培养基(CA)培养,培养后能够产生荧光的菌株被认为是OTA产生菌。同时分别用高效液相色谱法(HPLC)和OTA胶体金试纸条对所有菌株培养物中的OTA进行检测。【结果】以HPLC检测结果为标准,用紫外荧光法对OTA产生菌初筛的假阳性率较高,假阴性率为0;胶体金免疫试纸条法用于筛选OTA产生菌的假阴性率为47.4%,假阳性率为0,适合于产毒量高的菌种筛选。【结论】用紫外荧光法作为OTA产生菌的初筛方法可以排除大部分非OTA产生菌,漏检率低,从而大大降低HPLC检测的工作强度和检测成本;将紫外荧光法和HPLC定量分析结合使用,可以快速、准确地分离到OTA产生菌;将紫外荧光法和胶体金免疫试纸条相结合,可以快速筛选OTA产量高的菌株,且该方法较为经济,对环境和人体的污染较小。
[Objective] This paper aimed at screening out a method that can quickly identify the ochra- toxin-producing ability of the isolates and set foundation for ochratoxin detection and prevention. [Method] Three different methods of UV fluorescence,high performance liquid chromatography （HPLC） and colloi- dal gold immunoassay strip were used to screen out ochratoxin （OTA）-producer from 189 strains isolated from grape. In the UV fluorescence method,isolates belonging to Aspergillus section Nigri were cultivated in cococut cream medium （CCA） and other fungi in Czapek medium （CA）, respectively. The strains that produce fluorescent colour were supposed to be OTA-producer. HPLC and OTA colloidal gold test strip were also used to test OTA production in the culture at the same time. [Result] According to the HPLC results, the UV fluorescence method yielded high false-positive rate but no false-negative rate. The colloidal gold test strip method yielded false-negative rate of 47.4% and false-positive rate of 0. [Conclusion] UV fluorescence method could be used for primary screening for excluding most non-OTA-producing strains and thus significantly reduce the HPLC labour intensity and testing cost. Combination of the UV fluores- cence method and HPLC is a rapid and accurate method to screen out OTA-producing strains. Combination of the UV fluorescence and the colloidal gold test strip methods could be a good method to screen out thestrains that have high OTA-producing ability. Compared with HPLC method,this would be more conven- ient, cost lower and have lower contamination for environment and human.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2012年第3期169-174,共6页
Journal of Northwest A&F University(Natural Science Edition)
基金
农业部现代葡萄产业技术体系建设专项(nycytx-30-ch-03)
关键词
赭曲霉毒素A
紫外荧光法
OTA胶体金试纸条
产生菌筛选
Ochratoxin A ultraviolet fluorescence
OTA colloidal gold test strip
OTA-producing strainscreening
