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[利用聚合酶链反应鉴定球孢子菌病病原体]

[Identification of the agents of coccidioidomycosis using polymerase chain reaction].

作者信息

Tkachenko G A, Grishina M A, Antonov V A, Savchenko S S, Zamaraev V S, Lesovoĭ V S, Lipnitskiĭ A V

出版信息

Mol Gen Mikrobiol Virusol. 2007(4):25-31.

Abstract

Two pairs of primers for diagnosis of coccidioidomycosis using the method of PCR were constructed. One pair was used for identification of the two species of Coccidioides (C. immitis and C. posadasil) on the basis of MBP-1 gene. The other pair was chosen on the basis of SOWgp82 gene, which encodes an immunodominant, spherule outer wall glycoprotein for detecting only C. posadasii. The used primers allowed the agents of coccidioidomycosis to be detected using PCR with high sensitivity and specificity. The effective method of isolation of fungus DNA from soil contaminated with arthroconidia of Coccidioides spp. was developed. It includes guanidinthiocyanate-phenol-chloroform deproteinization followed by DNA purification using nuclear sorption.

摘要

构建了两对用于采用聚合酶链反应(PCR)方法诊断球孢子菌病的引物。一对引物基于MBP - 1基因用于鉴定球孢子菌的两个种(粗球孢子菌和波萨达斯球孢子菌)。另一对引物基于SOWgp82基因选择,该基因编码一种免疫显性的球形体外壁糖蛋白,用于仅检测波萨达斯球孢子菌。所使用的引物使得能够通过PCR以高灵敏度和特异性检测球孢子菌病病原体。开发了从被球孢子菌属关节孢子污染的土壤中有效分离真菌DNA的方法。该方法包括异硫氰酸胍 - 苯酚 - 氯仿脱蛋白,随后使用核吸附进行DNA纯化。

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