Kit, method, and device for sampling oral microbiome

A method of collecting a sample from an oral cavity of an animal, the method comprising:

• (i) providing a kit including:
• (a) a sample collection device having a handle and a head opposite the handle, the head comprising a plurality of bristles extending over an outer surface of the head, the handle including a first end and a second end with a longitudinal axis defined therebetween, and wherein the sample collection device is configured to brush a part of the oral cavity of the animal and to collect the sample therefrom, wherein the plurality of bristles are configured to capture the sample between respective bristles, and
• (b) a dry container without a buffer therein for storing the sample collection device therein, the dry container including a bottom segment couplable with a top segment to wholly house the sample collection device therein, the dry container having an open position for receiving at least the head in the top segment, and a closed position for receiving the head and the handle therein, wherein the dry container defines an aperture disposed at the top segment adjacent to the head to permit communication between an inside of the dry container with an external environment, wherein at least a portion of the handle is disposed within the bottom segment, wherein the sample collection device having a sample captured thereon and disposed in the dry container with the aperture protects stability of the sample for up to 14 days and up to 40 degrees Celsius;
• (ii) brushing at least one of gums, a gum-line, teeth, a tongue, a roof of a mouth, and/or another part of the oral cavity of the animal with the sample collection device to collect the sample thereon and between respective bristles;
• and
• (iii) drying the head and storing the sample collection device in the dry container for a duration of 5 days up to 14 days and up to 40 degrees Celsius.

A method of detecting one or more bacteria in an oral microbiome of a canine, the method comprising:

• (i) providing a first kit including:
• (a) a first sample collection device of a first type having a handle and a head opposite the handle, the first sample collection device configured to brush at least one of gums, a gum-line, teeth, a tongue, a roof of a mouth, and/or another part of an oral cavity of a canine to collect a first sample, and
• (b) a first dry container for storing the first sample collection device therein while drying the head of the first sample collection device, the first dry container having an open position for receiving at least the head and a closed position, wherein the first dry container defines an aperture to permit communication between an inside of the first dry container with an external environment, wherein the first sample collection device having a sample captured thereon and disposed in the first dry container with the aperture protects stability of the sample for up to 14 days and up to 40 degrees Celsius;
• (ii) providing a second kit including:
• (a) a second sample collection device of a second type having a handle and a head opposite the handle, the second sample collection device configured to brush at least one of gums, a gum-line, teeth, a tongue, a roof of a mouth, and/or another part of the oral cavity of the canine to collect a second sample, and
• (b) a second dry container for storing the second sample collection device therein while drying the head of the second sample collection device, the second dry container having an open position for receiving at least the head and a closed position, wherein the second dry container defines an aperture to permit communication between an inside of the second dry container with the external environment, wherein the second sample collection device having a sample captured thereon and disposed in the second dry container with the aperture protects stability of the sample for up to 14 days and up to 40 degrees Celsius;
• (iii) collecting the first sample using the first sample collection device by brushing at least one of gums, a gum-line, teeth, a tongue, a roof of a mouth, and/or another part of the oral cavity of the canine,
• (a) storing the first sample collection device within the first dry container for 5 days up to 14 days;
• (iv) collecting the second sample using the second sample collection device by brushing at least one of gums, a gum-line, teeth, a tongue, a roof of a mouth, and/or another part of the oral cavity of the canine,
• (a) storing the second sample collection device within the second dry container for 5 days up to 14 days;
• (v) performing a first assay on the first sample to measure a first value, the first value being an amount of a microbial nucleic acid to detect the one or more bacteria and/or to measure an amount of the one or more bacteria in the first sample;
• (vi) performing a second assay on the second sample to measure a second value, the second value being an amount of a microbial nucleic acid to detect the one or more bacteria and/or to measure an amount of the one or more bacteria in the second sample;
• wherein the first assay and the second assay are a qPCR assay against a 16S rRNA gene of the one or more bacteria selected from a group consisting of Peptostreptococcus sp., Synergistes sp., Clostridiales sp., Eubacterium nodatum, Selenomonas sp., Bacteroidetes sp., Odoribacter denticanis, Desulfomicrobium ovale, Moraxella sp., Bacteroides denticanoris, Fillifactor villosus, Porphyromonas canoris, Porphyromonas gulae, Treponema denticola, and Porphyromonas salivosa;
• (vii) comparing the first value and the second value to determine a relative abundance of Peptostreptococcaceae sp. in canine oral microbiome for each of the first sample and second sample;
• and
• (viii) diagnosing a likelihood of the canine having an oral disease or disorder based on the abundance of the Peptostreptococcaceae sp. in the oral microbiome of the canine;
• wherein the bacteria of the Peptostreptococcaceae sp. are selected from the group consisting of Peptostreptococcaceae XIII [G-1] sp., Peptostreptococcaceae COT-030, Peptostreptococcaceae COT-005/004, Peptostreptococcaceae COT-047, and/or Peptostreptococcaceae COT-019.