Methods for the diagnosis of bacterial vaginosis
View Patent ↗The present invention relates to methods for the diagnosis of bacterial vaginosis based on an analysis of a patient sample. For example, patient test samples are analyzed for the presence or absence of one or more lactobacilli and two or more pathogenic organisms. The presence or absence of one or more lactobacilli and two or more pathogenic organisms may be detected using PCR analysis of nucleic acid segments corresponding to each target organism. The quantity of the target organisms can then be used to determine a score which is indicative of a diagnosis of bacterial vaginosis.
1. A method for identifying bacterial vaginosis in a human female subject, the method comprising:
(a) obtaining a vaginal swab sample from the subject;
(b) measuring levels of Atopobium vaginae, Megasphaera genus and one or more Lactobacilli species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus crispatus , and Lactobacillus jensenii in the sample, wherein the levels of Atopobium vaginae, Megasphaera genus and of one or more Lactobacilli species are detected with one or more oligonucleotides capable of specifically hybridizing to a target nucleic acid sequence from Atopobium vaginae, Megasphaera genus, Lactobacillus acidophilus, Lactobacillus crispatus , and/or Lactobacillus jensenii , wherein levels of Gardnerella vaginalis and Mobiluncus sp are not measured; wherein the levels are determined with one or more oligonucleotide primers selected from SEQ ID NOs: 1, 2, 4, 5, 11, 12, 14 and 15, and complements thereof;
(c) calculating a diagnostic score as the ratio of a logarithmic function of the levels of the one or more Lactobacilli species and a logarithmic function of the levels of the Atopobium vaginae and Megasphaera genus; and
(d) identifying the subject as having bacterial vaginosis with the diagnostic score lower than a reference score.
2. A method for identifying bacterial vaginosis in a human female subject, the method comprising:
(a) obtaining a vaginal swab sample from the subject;
(b) measuring levels of Atopobium vaginae, Megasphaera genus and one or more Lactobacilli species selected from the group consisting of Lactobacillus acidophilus, Lactobacillus crispatus, and Lactobacillus jensenii in the sample, wherein the levels of Atopobium vaginae, Megasphaera genus and of one or more Lactobacilli species are detected with one or more oligonucleotides capable of specifically hybridizing to a target nucleic acid sequence from Atopobium vaginae, Megasphaera genus, Lactobacillus acidophilus, Lactobacillus crispalus , and/or Lactobacillus jensenii , wherein levels of Gardnerella vaginalis and Mobiluncus sp are not measured; wherein the levels are determined by PCR with oligonucleotide primer pairs comprising SEQ ID NOs: 1, 2, 4, 5, 11, 12, 14 and 15;
(c) calculating a diagnostic score as the ratio of a logarithmic function of the levels of the one or more Lactobacilli species and a logarithmic function of the levels of the Atopobium vaginae and Megasphaera genus; and
(d) identifying the subject as having bacterial vaginosis with the diagnostic score lower than a reference score.