| Literature DB >> 20728580 |
Mohammad Mehdi Feizabadi1, Araz Majnooni, Bizhan Nomanpour, Bahram Fatolahzadeh, Nafiseh Raji, Somayeh Delfani, Minoo Habibi, Soroor Asadi, Mahmood Parvin.
Abstract
Using oprL sequences, a TaqMan real time PCR was developed and used for quantitative detection of Pseudomonas aeruginosa from 99 broncoalveolar lavage and 11 sputum specimens collected from patients with health care associated pneumonia. All specimens were cultured on appropriate media to isolate bacteria. Twenty five specimens were positive by both methods. Polymicrobial infections were found in 13 specimens. Amplification of oprL in serial dilutions ranged from 10(9)CFU/ml to 10(2)CFU/ml. Standard curve of duplicated every dilution had slope 3.25±0.1 and R(2)>0.99 with SD 0.1. Our real time PCR assay showed high sensitivity (100%) and specificity (98.85%). This technique could detect and enumerate 100 bacteria directly from clinical specimens and showed that the threshold is 10(3)CFU/ml in cases with clinical symptoms. Our method can be used for quantitative detection of P. aeruginosa from BAL and sputum specimens in 1h and 10min.Entities:
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Year: 2010 PMID： 20728580 DOI： 10.1016/j.meegid.2010.08.008
Source DB: PubMed Journal: Infect Genet Evol ISSN： 1567-1348 Impact factor: 3.342