实时荧光PCR法对器官移植术后侵袭性肺曲霉病的早期诊断价值

The value of real-time PCR analysis of serum samples for early diagnosis of invasive pulmonary aspergillosis in organ translant recipients

目的评价实时荧光PCR检测血清曲霉DNA对器官移植术后侵袭性肺曲霉病的早期诊断价值。方法收集2004年1月至2006年3月在上海交通大学附属第一人民医院器官移植中心接受器官（或组织）移植伴肺部病变的器官移植术后59例患者的274份血清。按照欧洲癌症研究治疗组织及真菌研究组（EORTC／MSG）的标准分为侵袭性肺曲霉病（IPA）确诊、临床诊断、拟诊及非IPA患者，用实时荧光PCR法检测血清曲霉DNA。结果59例中男43例，女16例，平均年龄51．7岁。59例中确诊5例，临床诊断6例，拟诊18例，非IPA患者30例。确诊及临床诊断的11例中7例实时荧光PCR检测阳性；30例非IPA患者中2例实时荧光PCR检测阳性，考虑为假阳性。与痰培养、胸部CT及血清半乳甘露聚糖抗原（galactomannan，GM）检测比较，实时荧光PCR法出现阳性结果的时间较痰培养平均提前7．9d，较胸部CT平均提前6．0d，较GM检测平均提前1．5d。结论实时荧光PCR法对侵袭性肺曲霉病具有早期诊断意义。

Objective To study the value of AspergiUus real-time PCR as a tool for diagnosing invasive pulmonary aspergillosis（IPA） from serum samples in organ transplant recipients. Methods From Jan. 2004 to Mar. 2006, 59 organ translant recipients from Shanghai First People＇ s Hospital with high possibility of invasive pulmonary aspergillosis were evaluated. Of the 59 patients, 5 had proven, 6 had probable, and 18 had possible invasive Aspergillus infection according to European Organization for Research and Treatment of Cancer/Mycosis Study Group definitions. A total of 274 serum samples were analyzed using real-time PCR. Results Seven of these immunocompromised patients were PCR positive, correlating with positive sputum cultures, positive histology, and positive signs of CT scans. False negative results were found in 2 patients. They had positive PCR results without any correlation to clinical or other diagnostic criteria. Twenty-eight immunocompromised patients with a negative PCR result showed no evidence of invasive fungal disease. Compared with sputum culture, CT scan and galactomannan test, PCR positivity preceded diagnosis by a mean of 7.9 d, 6. 0 d, 1.5 d, respectively. Conclusion Aspergillus real-time PCR is a helpful test for the early diagnosis of IPA in high-risk organ transplant recipients.