Methods for diagnosing irritable bowel syndrome
View Patent ↗The invention provides an ELISA assay for the determination of serum mast cell β-tryptase levels using rabbit anti-tryptase as the capture antibody and alkaline phosphatase conjugated G3 as the detecting antibody. Luminescent substrate CPSD was used to enhance the assay sensitivity. Also provided are methods for aiding in the diagnosis of irritable bowel syndrome by detecting the serum level of β-tryptase, histamine and/or prostaglandin E 2 .
1. A method for aiding in the diagnosis of irritable bowel syndrome (IBS) in a subject, said method comprising:
(a) contacting a blood or serum sample from the subject with a β-tryptase binding moiety under conditions suitable to transform β-tryptase present in the sample into a complex comprising β-tryptase and the β-tryptase binding moiety; and
(b) determining the level of said complex, thereby determining the level of β-tryptase present in the sample.
2. The method of claim 1 , wherein the method further comprises:
(c) comparing the level of β-tryptase present in the sample to a control level, wherein a difference in the level of β-tryptase present in the sample relative to the control level is indicative of an increased likelihood of said subject having IBS.
3. The method of claim 2 , wherein the control level is the level of β-tryptase present in a blood or serum sample from a healthy subject.
4. The method of claim 3 , wherein an increased level of β-tryptase present in the sample relative to the control level is indicative of an increased likelihood of said subject having IBS.
5. The method of claim 3 , wherein the same or a reduced level of β-tryptase present in the sample relative to the control level is indicative of an increased likelihood of said subject not having IBS.
6. The method of claim 2 , wherein the control level is the level of β-tryptase present in a blood or serum sample from a subject with IBS.
7. The method of claim 6 , wherein the same or an increased level of β-tryptase present in the sample relative to the control level is indicative of an increased likelihood of said subject having IBS.
8. The method of claim 6 , wherein a reduced level of β-tryptase present in the sample relative to the control level is indicative of an increased likelihood of said subject not having IBS.
9. The method of claim 1 , wherein the method further comprises determining the level of histamine and/or prostaglandin E 2 (PGE 2 ) present in the sample.
10. A method for monitoring the progression or regression of irritable bowel syndrome (IBS) in a subject, the method comprising:
(a) contacting a first blood or serum sample taken from the subject at a first time with a β-tryptase binding moiety under conditions suitable to transform β-tryptase present in the sample into a complex comprising β-tryptase and the β-tryptase binding moiety;
(b) determining the level of said complex, thereby determining the level of β-tryptase present in the first sample;
(c) contacting a second blood or serum sample taken from the subject at a second time with a β-tryptase binding moiety under conditions suitable to transform β-tryptase present in the sample into a complex comprising β-tryptase and the β-tryptase binding moiety;
(d) determining the level of said complex, thereby determining the level of β-tryptase present in the second sample; and
(e) comparing the level of β-tryptase present in the first sample to the level of β-tryptase present in the second sample,
wherein a higher level of β-tryptase in the second sample relative to the first sample is indicative of the progression of IBS in the subject and a lower level of β-tryptase in the second sample relative to the first sample is indicative of the regression of IBS in the subject.