Detection and quantification of microorganisms using amplification and restriction enzyme analysis

A method of determining the relative ratios of microorganisms present in a microbiological flora, said method comprising:

• providing a sample of said microbiological flora;
• amplifying nucleic acid from the more than one species of microorganism present in said sample to produce an amplificate;
• subjecting the amplificate to at least one restriction digestion to produce restriction fragments from the more than one species of microorganism;
• preparing a resulting pattern based on the restriction fragments of the more than one species of microorganism in the sample;
• and
• analyzing the resulting pattern of restriction fragments to determine the relative ratios of microorganisms present in said flora.

A method of analyzing the composition of a bacterial flora, comprising:

• obtaining a sample of bacterial flora taken from the digestive tract of a subject;
• amplifying nucleic acid from more than one species of bacteria present in said sample with at least one primer of about 20 to 30 nucleotides directed to a nucleic acid believed to be present in said sample to produce an amplificate;
• subjecting the amplificate to restriction digestion;
• analyzing a resulting pattern of restriction fragments by comparing the restriction pattern of the sample with at least one reference restriction pattern;
• and determining relative abundance and relative ratios of said bacterial flora.

A method of analyzing a sample of microbiological flora containing more than one species of microorganism, said method comprising:

• providing the sample of said microbiological flora;
• subjecting the sample to at least one restriction digestion to produce restriction fragments from the more than one species of microorganism to produce a restriction digestion product;
• amplifying the restriction digestion product to produce an amplificate;
• and
• preparing a resulting pattern based on the restriction fragments of the more than one species of microorganism in the sample.