真菌性角膜溃疡的PCR检测

Detection of fungal corneal ulcer by polymerase chain reaction

目的：探讨临床拟诊为真菌性角膜溃疡PCR（polymerase chain reaction）快速检测的方法。方法：以源于医学条件致病性真菌18s rRNA基因保守区的一对寡核苷酸序列为通用引物，对临床拟诊为真菌性角膜溃疡的标本进行PCR检测．并与培养方法进行对比。结果：在400bp处出现DNA扩增带者为阳性。36份临床标本的真菌培养阳性率为58．33％。而经PCR扩增阳性率为86．11％。结论：真菌通用引物进行PCR反应检测真菌性角膜溃疡速度快、阳性率高，有助于真菌性角膜溃疡的快速诊断。

Objective： To find a method for the rapid detection of clinical suspect fungal corneal ulcer by polymerase chain reaction. Methods： A pair of oligonucleotide sequences,which was bases on the conserved region of 18s rRNA shared by medically important fungal corneal ulcer, was used as the general primers to amplify the DNAs from clinical suspect fungal corneal ulcer in a PCR assay, and the result was contrasted with culture. Resuits：A 400 bp specific DNA product was successfully amplified.The positive rate of fungi culture to 36 clinical specimens was 58.33%, and the positive rate of PCR amplification was 86.11%. Conclusion： PCR with the general primers is benefit for rapid detection of fungal corneal ulcer as quick speed and high positive rate.