Strain-dependent variation in 18S ribosomal DNA Copy numbers in Aspergillus fumigatus.

Enumerating Aspergillus fumigatus CFU can be challenging since CFU determination by plate count can be difficult. CFU determination by quantitative real-time PCR (qPCR), however, is becoming increasingly common and usually relies on detecting one of the subunits of the multicopy rRNA genes. This study was undertaken to determine if ribosomal DNA (rDNA) copy number was constant or variable among different A. fumigatus isolates. FKS1 was used as a single-copy control gene and was validated against single-copy (pyrG and ARG4) and multicopy (arsC) controls. The copy numbers of the 18S rDNA subunit were then determined for a variety of isolates and were found to vary with the strain, from 38 to 91 copies per genome. Investigation of the stability of the 18S rDNA copy number after exposure to a number of different environmental and growth conditions revealed that the copy number was stable, varying less than one copy across all conditions, including in isolates recovered from an animal model. These results suggest that while the ribosomal genes are excellent targets for enumeration by qPCR, the copy number should be determined prior to using them as targets for quantitative analysis.

Factors associated with acquisition of, or persistent colonization by, vaginal lactobacilli: role of hydrogen peroxide production.

To identify factors that predict sustained colonization by vaginal lactobacilli, microbiologic, behavioral, and demographic data were obtained from 101 nonpregnant women at baseline and at 4 and 8 months. A total of 272 isolates of lactobacilli were identified to the species level by use of whole chromosomal DNA homology to type strains. The predominant lactobacilli were the species Lactobacillus crispatus (38%) and L. jensenii (41%). Of 57 women initially colonized by H(2)O(2)-producing L. crispatus or L. jensenii, 23 (40%) remained colonized over 8 months, compared with 1 (5%) of 21 women colonized by other H(2)O(2)-producing species or by H(2)O(2)-negative strains (P=.01). Frequency of sexual intercourse (> or =1 sex act per week) was associated with loss of colonization with H(2)O(2)-producing lactobacilli (P=.018), as was antibiotic use (P< or =.0001). Other behavioral and demographic characteristics did not predict sustained colonization. The production of H(2)O(2) is closely linked with species and is a predictor for sustained long-term colonization of the vagina.