Next to the above given overview of hypotheses for the30 ontogenesis of depression and compounds that play a role in there, for someof these markers the relevance with respect to involvement and possiblydiagnosis of depression has been presented in the literature. A large, but far from complete list of the relevant literature is appended hereinafter. Further, the rationale for using ไลโปคาลิน-2 as marker can be5 derived from co-pending application NL 2007112.As indicated above, the markers can be used to affirm a diagnosis based on psychological and behavioral criteria. For each of the markers it can be indicated whether or not the level measured is indicative of thepresence of a mood disorder, such as depression. Information to that respect 10 can be found in the cited literature and the practical application of this knowledge is demonstrated in the experimental part of the present application Obviously, the more markers are used in the panel, and consequently, the more markers that are tested with a level that would suspect disease, the more reliable the diagnosis of depression can be made.15 Of course, if all or nearly all markers of the set that has been usedrespond positive for depression, the diagnosis should be considered definitive. Alternatively, a diagnostic score can be calculated as is exemplified on pages 8 and 9 of WO 2009/111595.20 On basis of the initial results with the markers, as presented inthe experimental section below, it has been established that a preferredembodiment of a panel of markers for serum/plasma assays which may be used for a diagnosis of a mood disorder, preferably depression, with a high confidentiality, ประกอบรวมด้วย the markers TNF-R2, คอร์ติซอล, ทรอมบอกเซน, 25 เอนโดธีลิน, เลปติน and vitamin D. In order to further increase the confidentiality of the diagnosis, this set may be extended with the group of biomarkers consisting of แคลโปรเทคติน, cAM, โซนูลิน and substance P. For a still further increase of the confidentiality the assay may be further extended with the markers BDNF, มิดไคน์, ไนโตรไทโรซีน, LTB4, 30 นิวโรเปปไทด์ Y, เทโลเมอเรส and อัลโดสเทอโรน.Similarly, for testing in urine only a set of markers whichprovides a reliable diagnosis consists of cGMP, คอร์ติซอล, แคลโปรเทคติน,ทรอมบอกเซน, อัลโดสเทอโรน, HVEM and substance P. A further improvement in the confidentiality of the diagnosis can be achieved to further include the 5 biomarkers เลปติน, LTB4, ไอโซโพรสเทน and มิดไคน์. An even further increase of the correctness of the diagnosis can be achieved by further including the
biomarkers cAMP, เอนโดธีลิน, TNF-R2 and นิวโรเปปไทด์ Y.
When mixing both serum/plasma biomarkers and urine
10 biomarkers, the minimal set of markers that is able to provide with a
reliable diagnosis (significance p<0.00001) consists of the biomarkers TNF-
R2 (s), คอร์ติซอล (s), ทรอมบอกเซน (s), เลปติน (s), เอนโดธีลิน (s), cGMP (u),
คอร์ติซอล (u), อัลโดสเทอโรน (u), ทรอมบอกเซน (u), HVEM (u) and substance P
(u), in which the suffix (s) or (u) indicates whether this marker should be 15 measured in serum/plasma (s) or in urine (u). In order to increase the
precision of the diagnosis, this set of biomarkers may be extended with the
group of Vitamin D (s), cAMP (s), โซนูลิน (s), substance P (s) and
แคลโปรเทคติน (u). A further increase in precision may be gained by further
combining the above mentioned marker sets with a further set of markers, 20 ซึ่งประกอบรวมด้วย แคลโปรเทคติน (s), เลปติน (u), LTB4 (u), ไอโซโพรสเทน (u) and มิดไคน์
(u).
The above-mentioned groups are also listed in Table 4 below.
25 While interpreting the results of such an assay, various
determinants such as sex, age, smoking status, urbanicity, food and alcohol intake should be taken into account. It can also be deduced from the cited literature (e.g. Bus et al., 2010) that the presence of a marker is related to a
factor such as sex, where in males a higher concentration is found than in
30 females, or to a factor such as age, where in elderly persons a higher level of
a marker is found than in younger persons. The measured values should of course be interpreted in the light of these correlations, which are known to the skilled person, as is demonstrated from the cited literature herein.
5 Preferably, the assay, when performed on urine, more preferably
on the first morning urine sample, also includes a simultaneous assay for
creatinine. Creatinine is one of the byproducts of protein metabolism. Under
normal conditions it is present in the blood and is excreted as a final
metabolite in the urine. Urine creatinine levels are routinely used as part of 10 kidney function diagnosis. In particular, altered creatinine levels in urine
are indicative of kidney diseases such as acute or chronic nephritis,
nephrosis, and the like. Because normative values for creatinine excretion
have been established, urine creatinine levels are also useful for correction
of assays for other compounds, as they document the adequacy of the urine 15 collection for such assays. In particular, the creatinine correction can be
used to correct for urine dilution, thus giving a possibility to standardize
measured concentrations irrespective of the water content of the urine
and/or the time of the day when the urine was produced. Further, changes
in renal function which influence rates of excretion, can be corrected by
20 measurement of creatinine in urine. Any values given in the experimental
part for assays done on urine have been corrected by measurement of
creatinine.
The biomarkers of the present การประดิษฐ์ or such a replacement
25 molecule are recognised by `biosensors', which may ประกอบรวมด้วย a ligand or
ligands capable of specific binding to the biomarker. Such biosensors are useful in detecting and/or quantifying the biomarker, preferably in
quantifying.
Especially useful biosensors are antibodies. The term 'antibody' as used herein may ประกอบรวมด้วย polyclonal, monoclonal, bispecific, humanised or chimeric antibodies, single chain antibodies, Fab fragments and F(ab')2
fragments, fragments produced by a Fab expression library, anti-idiotypic 5 antibodies and epitope-binding fragments. The term 'antibody' also refers to
immunoglobulin and T-cell รีเซปเตอร์ molecules, i.e. molecules that contain
an antigen-binding site that specifically binds an antigen. The
immunoglobulin molecules can be of any class (e.g. IgA, IgD, IgE, IgG and
IgM) or subclasses thereof. If the biomarker is a nucleic acid, a specific
10 probe can be used as `biosensor'. Preferably in such a case, the target DNA
is first amplified in a PCR reaction with suitable primer sequences.
The identification of the biomarkers that are specific for a mood
disorder, especially depression and more particularly major depressive
disorder is key to integration of diagnostic procedures and therapeutic
15 regimes. Appropriate diagnostic tools such as biosensors can be developed in
methods and uses of the การประดิษฐ์; and detection and quantification of the
biomarker can be performed using a biosensor in a microanalytical system,
a microengineered system, a microseparation system, an
immunochromatography system or other suitable analytical devices (such as 20 Raman or mass spectrography and the like). The biosensor may be
incorporated in an immunological method for detection of the biomarker(s),
or via electrical, thermal, magnetic, optical (e.g. hologram) or acoustic
technologies. Using these techniques, it is possible to detect the target
biomarker(s) at the anticipated concentrations found in biological samples. 25 Thus, according to a further aspect of the การประดิษฐ์ there is provided an
apparatus for diagnosing or monitoring a mood disorder, especially
depression and more particularly major depressive disorder which
ประกอบรวมด้วย one or more biosensors in a microanalytical, microengineered,
microseparation and/or immunochromatography system configured to detect 30 and/or quantify any of the biomarkers defined herein.
The biomarker(s) of the การประดิษฐ์ can be detected using a
biosensor incorporating technologies based on "smart" holograms, or high frequency acoustic systems, such systems are particularly amenable to "bar code" or array configurations.
5 In smart hologram sensors (Smart Holograms Ltd, Cambridge,
UK), a holographic image is stored in a thin polymer film that is sensitized to react specifically with the biomarker. On exposure, the biomarker reacts with the polymer leading to an alteration in the image displayed by the hologram.
10 The test result read-out can be a change in the optical brightness,
image, color and/or position of the image. For qualitative and semi-
quantitative applications, a sensor hologram can be read by eye, thus
removing the need for detection equipment. A simple color sensor can be
used to read the signal when quantitative measurements are required. 15 Opacity or color of the sample does not interfere with operation of the
sensor. The format of the sensor allows multiplexing for simultaneous
detection of several substances. Reversible and irreversible sensors can be
designed to meet different requirements, and continuous monitoring of a
particular biomarker of interest is feasible.
20 Suitably, methods for detection of one or more biomarkers
according to the การประดิษฐ์ combine biomolecular recognition with
appropriate means to convert detection of the presence or quantity of th
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