摘要
目的使临床系统性真菌病的诊断提高到分子水平。方法首先应用一对外侧的基于rDNA的真菌通用引物进行PCR扩增,此后应用新生隐球菌种特异性内侧引物对首轮扩增产物进行第二轮扩增鉴定。结果应用外侧引物对能对代表8属17种的25株医学重要真菌进行扩增,而对所有细菌和人DNA均为扩增阴性,内侧引物对仅对新生隐球菌获得扩增。应用两轮PCR扩增(通用PCR和随后的种特异性PCR)方法从获得临床标本到鉴定病原菌仅需8小时。
Objective The objective of the study was to develop the diagnosis of systemic mycoses to the sphere of molecular biology. Methods With the nested PCR methodology, a method for detecting fungal pathogens in clinical specimens was developed. The complete strategy consists of PCR amplification with the external fungal universal primers for rDNA and amplicon identification by the second cycle PCR amplification with the cryptococcus neoformans species specific primer pair. Results The external universal primers were capable of amplifying DNA from 25 strains representing 17 species (8 genera) of medically important fungi. Neither human nor a varity of pathogenic bacteria gave an amplified product. The internal species specific primers amplified DNA only from Cryptococcus neoformans. The use of these two PCRs in tandem allows the detection (universal PCR) and identification (species specific PCR) of a fungal pathogen within 8h from clinical specimens. Conclusion This PCR based detection and identification system provides a rapid laboratory method for the diagnosis of systemic mycoses and shows its potential for clinical applications.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
1998年第2期88-90,共3页
Chinese Journal of Infectious Diseases