Specific amplification of aspergillus fumigatus DNA by polymerase chain reation

ABSTRACT

Invasive aspergillosis [IA] is a life-threatening condition in immunocompromised patients. An early diagnosis is of great importance because early treatment may resolve this potentially fatal infection. Recently, the polymerase chain reaction [PCR] has been used successfully in detecting specific DNA of several pathogens. In this study, nested PCR was used to detect DNA specific for Aspergillus species isolated from bronchoalveolar lavage [BAL] fluid from patients with IA. In single PCR using the outer primers a specific 384-bp fragment was amplified. Similarly, by nested PCR with inner primers, a 357-bp fragment was amplified with DNA from Aspergillus fumigatus but not from the other microorganisms. The Southern blot hybridizations confirm the specificity of the PCR procedure for A.fumigatus using the cloned 374-bp PCR product probe. In conclusion, the nested PCR method appears to be quite rapid and specific