Evaluation of the Bacterial Vaginal Flora of 20 Postmenopausal Women by Direct (Nugent Score) and Molecular (Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis) Techniques

• Published in: The Journal of infectious diseases 2002-12, Vol.186 (12), p.1770-1780
• Main Authors: Burton, Jeremy P., Reid, Gregor
• Format: Article
• Language: English
• Subjects: Adult; Aged; Bacteria; Bacteria - genetics; Bacteria - isolation & purification; Bacterial vaginosis; Biological and medical sciences; Colony Count, Microbial; Demand side management; DNA Primers; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Female; Flora; Gels; Gentian Violet; Gynecology. Andrology. Obstetrics; Humans; Lactobacillus; Lactobacillus - isolation & purification; Major Article; Medical sciences; Microorganisms; Middle Aged; Phenazines; Polymerase Chain Reaction; Postmenopause; Puberal and climacteric disorders (male and female); Sequence Analysis, DNA; Vagina - microbiology; Vaginal Smears
• ISSN: 0022-1899; 1537-6613
• DOI: 10.1086/345761

To monitor the bacterial vaginal microflora of postmenopausal women, several approaches were used: (1) Nugent scoring of Gram-stained smears from vaginal swabs; (2) testing for bacteria, using specific polymerase chain reaction (PCR) primers; (3) PCR–denaturing gradient gel electrophoresis (DGGE) analysis of total bacterial DNA, amplified using eubacterial and Lactobacillus-specific primers; and (4) sequence analysis of amplified DNA fragments of interest from denaturing gradient gels. Of 20 women studied, 70% had either intermediate-grade bacterial colonization or bacterial vaginosis (BV), as defined by Nugent criteria, at the first sampling point. Subjects with normal Nugent scores had ⩾1 species of Lactobacillus detected by sequencing of dominant DNA fragments from denaturing gradient gels generated using eubacterial primers. Samples from women with Nugent scores that indicated BV had varied bacterial DGGE profiles, which emphasizes the diverse nature of such infections. This study indicates that asymptomatic BV appears to be much more common than is currently perceived; these findings may have implications for the health of postmenopausal women