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目的 探讨快速检测某些机会致病性真菌的方法。方法 以源于医学机会致病性真菌 1 8srRNA保守区的一对寡核苷酸序列为引物 ,利用聚合酶链反应对医学上重要机会致病性真菌DNA进行扩增。结果 3属 7种 2 9株重要机会致病性真菌 ,经扩增均出现一条 1 97bp的DNA片段 ,而对大肠埃希菌、金黄色葡萄球菌、绿脓假单胞菌、人白细胞则不扩增。以溴化乙啶染色 ,该PCR方法对白念珠菌DNA的最低检出限为 1 pg。 30份临床标本的真菌培养阳性率为2 3.3% ,而经扩增阳性率为 40 %。结论 该法具有较高的灵敏度与特异性 ,有可能应用于临床 ,以早期快速检测某些机会致病性真菌引起的深部真菌病
Objective To explore a method for rapid detection of some opportunistic fungi. Methods A pair of oligonucleotide primers derived from the 18srRNA conserved region of pathogenic fungi in medical science was used as a primer to amplify the medically important opportunistic pathogenic fungal DNA by polymerase chain reaction. Results Three genera and seven 29 kinds of pathogenic fungi were identified as important pathogenic fungi, and a DNA fragment of 1 97 bp appeared after amplification. However, for Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, human leukocytes were not Amplify. Ethidium bromide staining, the PCR method for Candida albicans DNA detection limit of 1 pg. The positive rate of fungal culture in 30 clinical specimens was 23.3%, while the positive rate was 40%. Conclusion The method has high sensitivity and specificity, it may be applied to clinical early rapid detection of some opportunistic fungi caused by fungal disease