title: Efficient method for mycobacterial DNA extraction in blood cultures aids rapid PCR identification of Mycobacterium tuberculosis and Mycobacterium avium.
abstract: The study presented here evaluated the utility of several methods of extracting mycobacterial nucleic acids from positive blood culture samples and examined the effect of each method on the performance of an in-house PCR used directly in the peripheral blood of 80 patients with AIDS to identify Mycobacterium spp. The modified Boom method for extracting DNA from blood cultures proved to be the most efficient, with subsequent PCR analysis yielding 100% positivity (7 samples positive for M. avium and 5 for M. tuberculosis). Only three of 12 patients with a positive blood culture had a PCR result positive for M. avium in peripheral blood. The identification of mycobacteria by PCR in blood culture took about 3 days, reducing the time to diagnosis by several weeks. These results demonstrate that PCR is a sensitive and quick method for identifying mycobacteria, especially when a good DNA extraction method is applied.
authors.affiliation: Laboratório Central do Estado, Ubaldino do Amaral, 545 Alto da XV, Curitiba-PR, Brazil.
authors.forename: S M
authors.initials: SM
authors.lastname: Nakatani
authors.forename: M
authors.initials: M
authors.lastname: Burger
authors.forename: M C
authors.initials: MC
authors.lastname: Assef
authors.forename: S R
authors.initials: SR
authors.lastname: Brockelt
authors.forename: L L
authors.initials: LL
authors.lastname: Cogo
authors.forename: I J T
authors.initials: IJ
authors.lastname: Messias-Reason
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000150:complications; D000293:Adolescent; D000328:Adult; D002648:Child; D004269:DNA, Bacterial / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009164:Mycobacterium Infections / Q000175:diagnosis; D015269:Mycobacterium avium Complex / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial