DETECTION OF ASPERGILLUS SPECIES DNA IN BRONCHOALVEOLAR LAVAGE SAMPLES BY COMPETITIVE PCR
DETECTION OF ASPERGILLUS SPECIES DNA IN BRONCHOALVEOLAR LAVAGE SAMPLES BY COMPETITIVE PCR
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DOI：
10.1128/jcm.33.5.1164-1168.1995
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发表时间：
1995-05-01
影响因子：
9.4
通讯作者：
FLEURYFEITH, J
中科院分区：
文献类型：
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作者：
BRETAGNE, S;COSTA, JM;FLEURYFEITH, J
建立了以支气管肺泡灌洗(BAL)标本诊断侵袭性肺曲霉菌病(IPA)的竞争性聚合酶链式反应方法。为此，我们测定了烟曲霉1kb线粒体DNA片段的序列。所用的引物允许烟曲霉、黄曲霉、黑曲霉和黄曲霉的DNA扩增，但不能扩增其他真菌和酵母的DNA。对55名连续入选的患者的BAL样本进行了测试。3个样本因内部竞争控制未能正确扩增而被排除。在28例免疫缺陷患者中，6例PCR阳性，3例死于IPA，BAL培养产生烟曲霉菌，3例培养阴性，未发生IPA。在15例人类免疫缺陷病毒阳性患者和9例免疫功能正常患者中，分别有5例和4例分别为PCR阳性和培养阴性，无一例发生曲霉病。因此，在3名患者中，聚合酶链式反应证实了IPA，但在没有发生曲霉病的患者中，有25%(12/49)的患者呈阳性结果。对于有曲霉病风险的患者，聚合酶链式反应阳性结果的预测价值似乎很低。此外，反应缓冲液或生物样品被分生曲霉污染的风险似乎很高，必须权衡作为常规程序的聚合酶链式反应检测的潜在诊断益处。
A competitive PCR assay involving the use of bronchoalveolar lavage (BAL) samples for the diagnosis of invasive pulmonary aspergillosis (IPA) was developed. For this purpose, a 1-kb mitochondrial DNA fragment of Aspergillus fumigatus was sequenced. The primers used allowed amplification of A. fumigatus, A. flavus, A. terreus, and A. niger DNAs but not DNAs of other fungi and yeasts. BAL samples from 55 consecutively enrolled patients were tested. Three samples were excluded because of failure of correct amplification of the internal competitive control. Of 28 immunocompromised patients, 6 were PCR positive; 3 died of IPA and their BAL cultures yielded A. fumigatus; and 3 were culture negative and did not develop IPA. Of 15 human immunodeficiency virus-positive patients and 9 immunocompetent patients, 5 and 4, respectively, were both PCR positive and culture negative, and none developed aspergillosis. Thus, PCR confirmed IPA in three patients but gave positive results for 25% (12 of 49) of the patients who did not develop aspergillosis. The predictive value of PCR-positive results seems low for patients at risk for aspergillosis. Moreover, the risk of contamination of reaction buffers or biological samples with Aspergillus conidia seems high and has to be weighed in regard to the potential diagnostic benefit of PCR testing as a routine procedure.