采用环介导等温扩增（LAMP）法检测巴西副球孢子菌的gp43

Detection of gp43 of Paracoccidioides brasiliensis by the loop-mediated isothermal amplification (LAMP) method.

作者信息

Endo Shigeo, Komori Takashi, Ricci Giannina, Sano Ayako, Yokoyama Koji, Ohori Akira, Kamei Katsuhiko, Franco Marcello, Miyaji Makoto, Nishimura Kazuko

机构信息

Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan.

出版信息

FEMS Microbiol Lett. 2004 May 1;234(1):93-7. doi: 10.1016/j.femsle.2004.03.015.

DOI:10.1016/j.femsle.2004.03.015

PMID:15109725

Abstract

Paracoccidioidomycosis is a deep mycosis caused by the thermo-dependent dimorphic fungus Paracoccidioides brasiliensis and is prevalent in Latin American countries. We detected the species specific gp43 gene of P. brasiliensis by loop-mediated isothermal amplification (LAMP) in 22 clinical and seven armadillo-derived isolates. The amplified DNA appeared as a ladder with a specific banding pattern. The advantage of the LAMP method is speed; only 3 h were necessary for identification of the organism and diagnosis of the disease. We were also able to obtain positive results from DNA extracted from a paraffin-embedded tissue sample of paracoccidioidomycosis, suggesting that this method may achieve clinical application in the near future.

摘要

副球孢子菌病是一种由温度依赖性双相真菌巴西副球孢子菌引起的深部真菌病，在拉丁美洲国家较为普遍。我们通过环介导等温扩增技术（LAMP）在22株临床分离株和7株犰狳来源的分离株中检测到巴西副球孢子菌的种特异性gp43基因。扩增的DNA呈现出具有特定条带模式的梯状条带。LAMP方法的优点是速度快；鉴定该病原体和诊断疾病仅需3小时。我们还能够从副球孢子菌病石蜡包埋组织样本中提取的DNA获得阳性结果，这表明该方法可能在不久的将来实现临床应用。