定量PCR检测对侵袭性真菌感染肺组织标本病原真菌的诊断价值

Value of quantitative polymerase chain reaction for diagnosis of invasive fungal infection in lung tissue samples

目的探讨应用定量聚合酶链反应（PCR）检测技术诊断侵袭性真菌感染肺组织标本病原真菌的临床意义。方法收集2010年6月至2016年6月间我院行手术切除且病理诊断为真菌感染的肺组织标本30例和同期非真菌感染的正常肺组织标本10例为研究对象，设计针对真菌的通用引物和针对隐球菌属和曲霉菌的特异性引物，采用PCR技术和双脱氧链终止法（Sanger法测序）检测石蜡包埋肺切除组织中的真菌。结果实验组33份真菌感染的肺组织标本PCR检测结果均为阳性，与病理诊断相符，Sanger测序可以鉴定真菌到种别。PCR检测与Sanger测序敏感度差异无统计学意义（100％比91．67％，P〉0．05），PCR检测与Sanger测序特异性差异无统计学意义（100％比100％，P〉0．05）；PCR检测种属鉴定率显著优于镜检（100％比81．82％，χ^2=6．60，P〈0．05），PCR检测与Sanger测序差异无统计学意义（100％比90．91％，χ^2=3．14，P〉0．05）。结论定量PCR检测用于诊断侵袭性真菌感染肺组织标本敏感性高、特异性好，具有一定参考意义。

Objective To investigate the clinical value of quantitative polymerase chain reaction （PCR） for diagnosis of invasive fungal infections in lung tissue samples. Methods 33 cases of confirmed fungal infection and 10 cases of confirmed non-fungal infections from June 2010 to June 2016 were selected. Universal primers for fungal and specific primers for Cryptococcus and Aspergillus were designed,and the method of PCR and Sanger sequencing were used. Results 33 cases of lung tissue specimens with fungal infection were all positive using PCR detection, which was consistent with pathological diagnosis. Sanger sequencing could identify fungi to the specific spaces. Sensitivity of PCR detection showed no statistically significant difference with that of Sanger sequencing （100% vs 91.67 %, P ）0.05）. Specificity of PCR detection showed no statistically significant difference with that of Sanger sequencing （100% vs 100%, P ）0.05）. The species identification rate of PCR detection was significantly better than that of microscopy （100% vs 81.82%, χ^2 = 6.60, P 〈0.05）, and was similar with that of Sanger sequencing （100% vs 90.91$, χ^2 = 3.14, P〈0.05）. Conclusions Quantitative PCR detection used in the diagnosis of invasive fungal infection lung tissue samples has certain reference significance with high sensitivity, and good specificity.