US 9,809,861 B2
Method of evaluating inhibitory effect on damp-dry malodor
Asako Mitani, Tokyo (JP); Hiromi Kubota, Tochigi (JP); Yu Niwano, Wakayama (JP); Kohei Takeuchi, Funabashi (JP); Atsushi Tanaka, Hannan (JP); and Noriko Yamaguchi, Wakayama (JP)
Assigned to Kao Corporation, Tokyo (JP)
Appl. No. 14/375,998
Filed by Kao Corporation, Chuo-ku, Tokyo (JP)
PCT Filed May 28, 2013, PCT No. PCT/JP2013/065306
§ 371(c)(1), (2) Date Jul. 31, 2014,
PCT Pub. No. WO2013/180302, PCT Pub. Date Dec. 5, 2013.
Claims priority of application No. 2012-122032 (JP), filed on May 29, 2012; and application No. 2013-109303 (JP), filed on May 23, 2013.
Prior Publication US 2015/0031041 A1, Jan. 29, 2015
Int. Cl. C12P 19/34 (2006.01); C12Q 1/68 (2006.01)
CPC C12Q 1/6897 (2013.01) [C12Q 1/689 (2013.01); C12Q 2600/136 (2013.01); C12Q 2600/158 (2013.01)] 35 Claims
 
1. A method of evaluating an inhibitory effect on damp-dry malodor, comprising the steps of:
bringing microorganisms which produce damp-dry malodor-causing substances and a test substance into contact with each other in the presence of a sebaceous dirt component;
detecting expression of at least one gene selected from the group consisting of a fatty acid desaturase gene and a β oxidation-related enzyme gene derived from the microorganisms; and thereby
evaluating a damp-dry malodor inhibitory function of the test substance based on a change in expression amount of the at least one gene;
wherein the expression of the at least one gene is detected by using at least one oligonucleotide pair selected from the group consisting of:
an oligonucleotide pair containing oligonucleotides (a) and (b) as described below,
an oligonucleotide pair containing oligonucleotides (c) and (d) as described below,
an oligonucleotide pair containing oligonucleotides (e) and (f) as described below,
an oligonucleotide pair containing oligonucleotides (g) and (h) as described below,
an oligonucleotide pair containing oligonucleotides (i) and (j) as described below,
an oligonucleotide pair containing oligonucleotides (k) and (l) as described below,
an oligonucleotide pair containing oligonucleotides (m) and (n) as described below,
an oligonucleotide pair containing oligonucleotides (o) and (p) as described below,
an oligonucleotide pair containing oligonucleotides (q) and (r) as described below,
an oligonucleotide pair containing oligonucleotides (s) and (t) as described below,
an oligonucleotide pair containing oligonucleotides (u) and (v) as described below,
an oligonucleotide pair containing oligonucleotides (w) and (x) as described below,
an oligonucleotide pair containing oligonucleotides (y) and (z) as described below,
an oligonucleotide pair containing oligonucleotides (a1) and (b1) as described below, and
an oligonucleotide pair containing oligonucleotides (c1) and (d1) as described below:
wherein
oligonucleotide (a) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 13, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 13 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (b) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 14, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 14 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (c) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 15, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 15 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (d) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 16, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 16 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (e) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 17, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 17 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (f) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 18, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 18 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (g) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 19, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 19 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (h) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 20, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 20 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (i) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 21, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 21 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (j) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 22, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 22 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (k) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 23, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 23 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (l) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 24, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 24 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the fatty acid desaturase gene;
oligonucleotide (m) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 25, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 25 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (n) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 26, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 26 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (o) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 27, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 27 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (p) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 28, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 28 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (q) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 29, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 29 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (r) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 30, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 30 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (s) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 31, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 31 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (t) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 32, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 32 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (u) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 33, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 33 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (v) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 34, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 34 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (w) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 35, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 35 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (x) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 36, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 36 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (y) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 37, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 37 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (z) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 38, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 38 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (a1) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 39, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 39 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (b1) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 40, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 40 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene;
oligonucleotide (c1) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 41, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 41 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene; and
oligonucleotide (d1) is an oligonucleotide having the base sequence set forth in SEQ ID NO: 42, or an oligonucleotide having a base sequence set forth in SEQ ID NO: 42 with a substitution, deletion, insertion, or addition of 1 to 3 nucleotides which can be used for detecting the β oxidation-related enzyme gene.