Method for diagnosis of and following a bacterial vaginosis by molecular quantification

A method for in vitro diagnosis and follow-up of the vaginal bacterial flora status in relation to the presence of bacterial vaginosis, and for monitoring its treatment where applicable, the method comprising:

• 1) quantifying the concentrations of the bacteria Atopobium vaginae and Gardnerella vaginalis by
• determining the concentrations of-the specific sequences of said bacteria Atopobium vaginae and Gardnerella vaginalis present in a single copy in the DNA of said bacteria Atopobium vaginae and Gardnerella vaginalis, and of a specific sequence of a human gene present in all biological specimens containing human cells, in the DNA extracted from a vaginal discharge specimen from a patient, said specific sequences being less than 150 nucleotides in size,
• enzymatically co-amplifying said specific sequences contained, on the one hand, in said DNA extracted from the specimen and, on the other hand, in samples of synthetic DNA fragments including each of said specific sequences of said bacteria and said specific sequence of a human gene present in all biological human cell specimens, said samples serving as calibration standards for quantifying the DNA,
• detecting and quantifying said amplified fragments with the aid of probes labeled with sequences different from those of the amplification primers for each of said specific sequences of said bacteria Atopobium vaginae and Gardnerella vaginalis and said specific sequence of a human gene present in all biological human cell specimens, and
• 2) determining the presence of bacterial vaginosis or failure of the on-going treatment if the concentrations of DNA fragments of said two specific sequences of the bacteria Atopobium vaginae and Gardnerella vaginalis in a specimen of the patient's vaginal discharge containing at least 104 human cells/mL are such that at least one of the following two conditions a) and b) is met:
• a) the concentration Ca of said DNA fragment of Atopobium vaginae is greater than or equal to 108 copies/mL, and
• b) the concentration Cg of said DNA fragment of Gardnerella vaginalis is greater than or equal to 109 copies/mL.