Detection and Identification of Aspergillus fumigatus in BAL Samples of Patients with Suspected Tuberculosis by Nested-PCR

Background and Aim: Aspergillus fumigatus is the cause of more than 90% of invasive pulmonary aspergillosis. The aim of this study is identification and detection of A.fumigatus in bronchoalveolar lavage (BAL) samples patients suspected to have tuberculosis by using of Nested-PCR.Materials and Methods: In this assay, a set of 100 BAL samples which were collected in 9 month, between the years 2012-2013, from patients with suspected tuberculosis in Tehran hospital, were examined by direct, culture and molecular tests. Manual phenol-chloroform method was used in order to extract DNA. Specific primers of Aspergillus genus and beta-actin primers were used for duplex-PCR. A pairs of internal primers were utilized in order to identification of A.fumigatus (Nested-PCR method). Finally, some of the PCR-products were sequenced for confirming results. Results: Overall, 12 samples (12%) were aspergillus-positive by molecular test while 11% and 10% samples were positive by direct and culture test, respectively. The sensitivities and specificities of PCR method compare with direct examination was 100% and 98.8%, respectively. The study also showed that sensitivities and specificities of PCR method compare with culture was 100% and 97.7%, respectively. After sequencing PCR product sample, isolated species were recognized as A. fumigatus. Conclusions: This study showed that PCR method in compare with the direct method and the culture has a higher sensitivity in the detection of Aspergillus species, also Present study displayed that can be consider Aspergillus species and A. fumigatus in suspicious to tuberculosis BAL samples.

Detection and Identification of Aspergillus fumigatus in BAL Samples of Patients with Suspected Tuberculosis by Nested-PCR Keywords:

Aspergillus fumigatus , Bronchoalveolar lavage (BAL) , PCR