Biomarkers for diagnosing schizophrenia and bipolar disorder

a biomarker and schizophrenia technology, applied in the field of identification and selection of novel biomarkers, can solve the problems of difficult diagnosis of schizophrenia, inconvenience for patients, and increased difficulty in diagnosing schizophrenia, and achieve the effect of monitoring the efficacy of treatmen

Inactive Publication Date: 2005-09-22
GENENEWS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009] The invention relates to the identification and selection of novel biomarkers and the identification and selection of novel biomarker combinations which are differentially expressed in blood and useful in diagnosing schizophrenia and/or bipolar disorder as well as monitoring therapeutic efficacy of treatment for schizophrenia and/or bipolar disorder. The measurement of expression levels of the products of the biomarkers and combinations of biomarkers of the invention can be used to diagnose schizophrenia and/or bipolar disorder. Measurement of the expression level of products of biomarkers of the invention using polynucleotides and prot

Problems solved by technology

Relying on symptomatic history makes diagnosis of schizophrenia difficult, particularly since no single symptom is definitive for diagnosis.
Diagnosis of schizophrenia is made even harder because it is often difficult to differentiate schizophrenia from other mental disorders including bipolar disorder, schizoaffective disorder, and brief psychotic disorder.
Although recently brain imaging techniques have been utilized as a tool towards diagnosis, this is costly, is inconvenient to patients, and is not considered very reliable.
Diagnosis can be difficult because the first episode of mania may go undetected,

Method used

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  • Biomarkers for diagnosing schizophrenia and bipolar disorder
  • Biomarkers for diagnosing schizophrenia and bipolar disorder
  • Biomarkers for diagnosing schizophrenia and bipolar disorder

Examples

Experimental program
Comparison scheme
Effect test

example 1

RNA Isolation from Lysed Blood

[0547] 10 ml whole blood is obtained in a Vacutainer and spun at 2,000 rpm for 5 min at 4° C. and the plasma layer removed. Lysis Buffer is added to blood sample in a ratio of 3 parts Lysis Buffer to 1 part blood (Lysis Buffer (IL) 0.6 g EDTA; 1.0 g KHCO2, 8.2 g NH4Cl adjusted to pH 7.4 (using NaOH)). Sample is mixed and placed on ice for 5-10 minutes until transparent. Lysed sample is centrifuged at 1000 rpm for 10 minutes at 4° C., and supernatant is aspirated. Pellet is resuspended in 5 ml Lysis Buffer, and centrifuged again at 1000 rpm for 10 minutes at 4° C. Pelleted cells are homogenized using TRIzol® (D (GIBCO / BRL) in a ratio of approximately 6 ml of TRIzol® for every 10 ml of the original blood sample and vortexed well. Samples are left for 5 minutes at room temperature. RNA is extracted using 1.2 ml of chloroform per 1 ml of TRIzol®. Sample is centrifuged at 12,000×g for 5 minutes at 4° C. and upper layer is collected. To upper layer, isoprop...

example 2

From Whole Blood

[0548] 100 ul whole blood is obtained in a microcentrifuge tube and spun at 2,000 rpm (800 g) for 5 min at 4° C. and the supernatant removed. Pelleted cells are homogenized using TRIzol (GIBCO / BRL) in a ratio of approximately 6 μl of TRIzol for every 10 μl of the original blood sample and vortexed well. Samples are left for 5 minutes at room temperature. RNA is extracted using 12 μl of chloroform per 10 μl of TRIzol. Sample is centrifuged at 12,000×g for 5 minutes at 4° C. and upper layer is collected. To upper layer, isopropanol is added in ratio of 5 μl per 10 μl of TRIzol. Sample is left overnight at −20° C. or for one hour at −20° C. RNA is pelleted in accordance with known methods, RNA pellet air dried, and pellet resuspended in DEPC treated ddH2O. RNA samples can also be stored in 75% ethanol where the samples are stable at room temperature for transportation.

From Centrifuged Lysed Blood

[0549] 10 ml whole blood is obtained in a Vacutainer and spun at 2,000...

example 3

Target Nucleic Acid Preparation and Hybridization

Preparation of Fluorescent DNA Probe from mRNA

[0551] Fluorescently labeled target nucleic acid samples of RNA are prepared for analysis with an array of the invention.

[0552] 1 μg Oligo-dT primers are annealed to 10 ug of total RNA isolated from blood from patient diagnosed with schizophrenia and / or bipolar disorder or suspected of having schizophrenia and / or bipolar disorder in a total volume of 10 ul, by heating to 70° C. for 10 min, and cooled on ice. The mRNA is reverse transcribed by incubating the sample at 42° C. for 40 min in a 25 ll volume containing a final concentration of 50 mM Tris-HCl (pH 8.3), 75 mM KCl, 3 mM MgC12, 25 mM DTT, 25 mM unlabeled dNTPs, 400 units of Superscript II (200 U / uL, Gibco BRL), and 15 mM of Cy3 or Cy5 (Amersham). The reaction is stopped by the addition of 2.5 μl of 55500 mM EDTA and 5 μl of 1M NaOH, and incubation at 65° C. for 10 min. The reaction mixture is neutralized by addition of 12.5 μl ...

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Abstract

The invention relates to the identification and selection of novel biomarkers and the identification and selection of novel biomarker combinations which are differentially expressed in blood and useful in diagnosing schizophrenia and/or bipolar disorder as well as monitoring therapeutic efficacy of treatment for schizophrenia or bipolar disorder. The measurement of expression levels of the products of the biomarkers and combinations of biomarkers of the invention can be used to diagnose schizophrenia and/or bipolar disorder. Measurement of the expression level of products of biomarkers of the invention using polynucleotides and proteins which specifically and/or selectively hybridize to the products of the biomarkers of the invention are also encompassed within the scope of the invention as are compositions and kits containing said polynucleotides and proteins. Further encompassed by the invention is the use of the polynucleotides and proteins to monitor the efficacy of therapeutic regimens. The invention also provides for the identification of methods of using the products of the biomarkers of the invention in the identification of novel therapeutic targets of schizophrenia and/or bipolar disorder and a method of screening the genes of said biomarkers for additional markers of disease.

Description

RELATED APPLICATIONS [0001] This application is a Continuation-in-Part of 10 / 812,731, filed Mar. 30, 2004 which is a continuation in part of 10 / 802,875, filed Mar. 12, 2004, which a continuation in part of application Ser. No. 10 / 601,518, filed on Jun. 20, 2003, which is a continuation-in-part of application Ser. No. 10 / 268,730 filed on Oct. 9, 2002, which is a continuation of U.S. application Ser. No. 09 / 477,148 filed Jan. 4, 2000, now abandoned, which claims the benefit of U.S. Provisional Application No. 60 / 115,125 filed on Jan. 6, 1999. Each of these applications is incorporated herein by reference in their entirety, including figures and drawings.1. FIELD OF THE INVENTION [0002] The invention relates to the identification and selection of novel biomarkers and the identification and selection of novel biomarker combinations which are differentially expressed in individuals with schizophrenia and / or bipolar disorder as well as a means of selecting the novel biomarker combinations...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12Q1/68
CPCC12Q1/6883G01N21/6486C12Q2600/158Y02A90/10
Inventor LIEW, CHOONG-CHINYAGER, THOMASDEMPSEY, ADAMCHAO, SAMUEL
Owner GENENEWS
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