PCR结合反向斑点杂交在侵袭性肺曲霉病早期诊断中的价值

The significance of PCR reverse dot blot hybridization in early diagnosis of invasive pulmonary aspergillosis

目的评价和比较血清半乳甘露聚糖(GM)浓度测定和PCR结合反向斑点杂交实验在侵袭性肺曲霉病(IPA)早期诊断中的临床价值。方法用酶联免疫吸附试验(ELISA)检测血清中GM的浓度;PCR结合反向斑点杂交法检测各组小鼠血清、肺泡灌洗液、肺组织中的曲霉菌DNA。结果 ELISA法测定GM浓度中,实验组血清、BALF的GM实验敏感性、特异性与对照组间差异有统计学意义(P<0.05);反向斑点杂交检测血清、肺泡灌洗液、肺组织中曲霉菌DNA含量,实验组阳性率明显高于对照组(P<0.05)。用血清标本的反向斑点杂交实验与GM实验、组织病理阳性率进行比较,组织病理的阳性率最高,其次是反向斑点杂交。三种方法的差别均有统计学意义(P<0.05)。结论 GM-ELI-SA检测是一项较灵敏、快速的诊断方法,反向斑点杂交实验在IPA早期诊断上的作用优于GM实验。

Objective To evaluate and compare the clinical value of the serum galactomannan （GM）detection and PC reverse dot blot hybridization experiments in the early diagnosis of invasive pulmonary aspergillosis （IPA）. Methods Enzyme-linked immunosorbent assay （ELISA） was used to detect the galactomannan （GM） concentration in the serum and bronchoalveolar lavage fluid （BALF） of rats. PCR combined with reverse dot blot hybridization was used to detect the aspergillus DNA in the mice serum,bronchoalveolar lavage fluid and lung tissue. Results In detection of GM using ELISA, the differences of sensitivity and specificity between experiment group and control group were significantly different （P 〈0.05）. In detection of the aspergillus DNA using PCR combined with reverse dot blot hybridization,the positive rate of the experimental group was significantly higher than that of the control group,and the difference was statistically significant （P 〈 0.05）.In comparison with histopathology and GM test,the highest positive rate was observed in histopathology method, followed by reverse dot blot hybridization, the difference of positive rates in the three methods was statistically significant （P 〈0.05）. Conclusion GM-ELISA test was a quite sensitive and rapid method in diagnosis of IPA. While the reverse dot blot hybridization was superior to GM test in the early diagnosis of IPA.