病原性真菌PCR检测方法的建立

目的：建立一种快速、灵敏和特异的病原性真菌的检测方法。方法：选取真菌18S rRNA保守区的一对寡核苷酸序列作为通用引物，对10种真菌、3种常见细菌及80例临床标本进行PCR扩增。结果：所试10种真菌均能扩增出一条约400bp的DNA片段，而3种细菌则无此扩增条带。通过对80例临床标本的检测，证实PCR法和常规培养法结果基本一致。此外，对白色念珠菌检测的最低限为10pg的DNA。结论：PCR方法检测临床常见真菌有较高的灵敏性与特异性，有助于临床真菌感染性疾病的诊断与治疗。

Detection and identification of medical important fungi by PCR

Aim: To develop a method with high sensitivity, specialty and rapidity to detect clinical important fungi by PCR. Methods: A pair of oligonucleotide sequences, which were selected from the conserved region of 18s rRNA shared by important medical fungi, were used as the primers to amplify the DNA of 10 species of fungi, 3 species of bacteria and 80 clinical samples. Results: A 400 bp specific DNA product emerged from 10 species of fungi, but not from 3 species of bacteria. The results obtained from PCR and general culture isolates were almost the same. The lower limit of detection of this PCR assay was 10 pg of Candida albicans genomic DNA. Conclusion: The PCR method is proved to be a fast, sensitive and specific technology to detect clinical fungi, and is helpful to the diagnoses and treatment of the diseases caused by fungi.