摘要
目的建立新的马拉色菌菌种鉴定方法,提高菌种的检出率,避免培养造成的误差,适合大样本量流行病学调查。方法采用无菌敷贴采集正常皮肤或皮损区标本,1%Triton X-100洗涤液反复洗涤后进行DNA提取,设计马拉色菌种特异性引物进行巢式PCR扩增,并以标准株作为对照,分离鉴定菌种构成。结果对10个种的标准株,特异引物均能扩出特异性条带。以标准株特异条带为参照,从15例正常志愿者头面部和10例花斑糠疹患者皮损区分离标本中均扩增出特异条带,显示构成以球形及限制马拉色菌为主(分别为34.30%,21.90%和34.50%,27.60%)。结论利用非培养直接DNA提取法能从正常皮肤及皮损区鉴定马拉色菌菌种构成。
Objective To find an approach for improving isolating rate, avoiding error of identifying Malassezia species, and finally providing benefit for the large sample epidemiological investigation. Methods Cutaneous micro- organisms were collected by applying transparent dressing and thereafter washed by 1% Triton X-100 solu- tion. DNA was extracted directly from the eluant, followed by nested PCR using designed specie-specific primers. Type species were chosen as control. Results Using species-specific primers, all 10 types of species showed specific bands in agarose gel. According to these specific bands of specie types, clinical strains isola- ted from 15 normal volunteers and 10 pityriasis versicolor patients also showed specific bands. In particular, M. globosa（34.30% or 34.50% ） and M. restricta（ 21. 90% or 27.60% ） were predominate species in these cases. Conclusion Non-culture method based on nested PCR was a direct assay for identification of Malas- sezia from lesional or non-lesional skin.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2014年第3期242-247,共6页
The Chinese Journal of Dermatovenereology
基金
国家自然科学基金青年基金(31100020)
四川省卫生厅科研课题(100141)
川北医学院苗圃基金(MP-ZK-34)
