Method of preparing a standard diagnostic gene transcript pattern

Abstract

A method for preparing a gene transcript pattern probe kit characteristic of a disease or condition or a stage thereof in a prokaryotic or eukaryotic organism using mRNA which is differentially expressed in the disease or condition or stage as probes, methods of diagnosis using the method and kits for performing the same are disclosed.

1 - 17 . (canceled) 
     
     
         18 . A method of obtaining isolated selected mRNA species or isolated selected cDNA species useful for diagnosing or identifying schizophrenia or a stage thereof in a human comprising the steps of:
 (a) isolating mRNA from cells from blood of more than one human who are known to have schizophrenia or a stage thereof (schizophrenia sample), wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA;   (b) isolating mRNA from corresponding cells from blood of more than one corresponding normal human (normal sample), wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA;   (c) separating mRNA species or cDNA species present within each of the resulting isolated mRNA or isolated cDNA of step (a) and (b), wherein the resulting separated mRNA species are optionally subject to reverse transcription to obtain separated cDNA species;   (d) selecting two or more mRNA species or two or more cDNA species from the resulting separated mRNA species or resulting separated cDNA species obtained in step (c), respectively, which are present at a different level in the normal sample than in the schizophrenia sample by identifying a signal corresponding to each mRNA species or cDNA species, wherein the resulting selected two or more mRNA species are optionally subjected to reverse transcription to obtain two or more selected cDNA species; and   (e) isolating the resulting two or more selected mRNA species or resulting two or more selected cDNA species obtained in step (d) to obtain isolated selected mRNA species or isolated selected cDNA species, wherein the resulting isolated selected mRNA species are optionally subjected to reverse transcription to obtain isolated selected cDNA species.   
     
     
         19 . A method of preparing a gene transcript pattern probe kit comprising the steps of:
 (a) isolating mRNA from cells from blood of more than one human who are known to have schizophrenia or a stage thereof (schizophrenia sample), wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA;   (b) isolating mRNA from corresponding cells from blood of more than one corresponding normal humans (normal sample), wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA;   (c) separating mRNA species or cDNA species present within each of the resulting isolated mRNA or isolated cDNA of step (a) and (b), wherein the resulting separated mRNA species are optionally subject to reverse transcription to obtain separated cDNA species;   (d) selecting two or more mRNA species or two or more cDNA species from the resulting separated mRNA species or resulting separated cDNA species obtained in step (c), respectively, which are present at a different level in the normal sample than in the schizophrenia sample by identifying a signal corresponding to each mRNA species or cDNA species, wherein the resulting selected two or more mRNA species are optionally subjected to reverse transcription to obtain two or more selected cDNA species;   (e) isolating the resulting two or more selected mRNA species or resulting two or more selected cDNA species obtained in step (d) to obtain isolated selected mRNA species or isolated selected cDNA species, wherein the resulting isolated selected mRNA species are optionally subjected to reverse transcription to obtain isolated selected cDNA species; and   (f) preparing at least one solid support carrying the resulting isolated selected mRNA species or isolated selected cDNA species of step (e) so as to form a gene transcript pattern probe kit.   
     
     
         20 . The method as claimed in  claim 19 , wherein said separation in step (c) is performed by a non-sequence based separation technique. 
     
     
         21 . The method as claimed in  claim 19 , wherein in steps (a) and (b), the resulting isolated mRNA is subjected to reverse transcription to obtain isolated cDNA. 
     
     
         22 . The method as claimed in  claim 21 , where said isolated cDNA is amplified. 
     
     
         23 . The method as claimed in  claim 19 , wherein in step (e), between 50 and 100 mRNA species or cDNA species are isolated and selected. 
     
     
         24 . The method as claimed in  claim 19 , wherein, in step (e), between 10 and 500 mRNA species or cDNA species are isolated and selected. 
     
     
         25 . The method as claimed in  claim 19 , wherein, in step (c), said separation technique is gel electrophoresis. 
     
     
         26 . The method as claimed in  claim 19 , wherein, prior to step (f), the resulting isolated selected mRNA species or isolated selected cDNA species of step (e) are amplified. 
     
     
         27 . A method of preparing a standard gene transcript pattern characteristic of schizophrenia or a stage thereof of a human comprising the steps of:
 (a) isolating mRNA from cells from blood of more than one human who are known to have schizophrenia or a stage thereof (schizophrenia sample), wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA,   (b) hybridizing the resulting isolated mRNA or isolated cDNA of step (a) to two or more mRNA species which are present at a different level in cells in a blood sample from more than one normal human than in corresponding cells in a blood sample from more than one humans known to have schizophrenia or a stage thereof, or to two or more cDNA species transcribed from said mRNA species, wherein the two or more mRNA species or cDNA species are specific for schizophrenia or a stage thereof, wherein said mRNA or cDNA species are carried on a solid support; and   (c) assessing the amount of hybridization so as to obtain said standard gene transcript pattern.   
     
     
         28 . A method of preparing a human test gene transcript pattern for schizophrenia or a stage thereof comprising the steps of:
 (a) isolating mRNA from cells from blood of a test human, wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA,   (b) hybridizing the resulting isolated mRNA or isolated cDNA of step (a) to two or more mRNA species which are present at a different level in cells in a blood sample from more then one normal human than in corresponding cells in a blood sample from more than one human known to have schizophrenia or a stage thereof, or to two or more cDNA species transcribed from said mRNA species, wherein the two or more mRNA species or cDNA species are specific for schizophrenia or a stage thereof, wherein said mRNA or cDNA species are carried on a solid support; and   (c) assessing the amount of hybridization so as to obtain said test gene transcript pattern.   
     
     
         29 . A method of diagnosing or identifying schizophrenia or a stage thereof in a test human comprising the steps of:
 (a) isolating mRNA from cells from blood of a test human, wherein the resulting isolated mRNA is optionally subjected to reverse transcription to obtain isolated cDNA;   (b) hybridizing the resulting isolated mRNA or isolated cDNA of step (a) to two or more mRNA species which are present at a different level in cells in a blood sample from more than one normal human than in corresponding cells in a blood sample from more than one human known to have schizophrenia or a stage thereof, or to two or more cDNA species transcribed from said mRNA species, wherein the two or more mRNA species or cDNA species are specific for schizophrenia or a stage thereof, wherein said mRNA or cDNA species are carried on a solid support;   (c) assessing the amount of hybridization so as to obtain a hybridization pattern; and   (d) comparing the resulting hybridization pattern obtained in step (c) with a hybridization pattern obtained by hybridizing isolated mRNA or isolated cDNA prepared from corresponding cells from blood of more than one corresponding human known to schizophrenia or a stage thereof to the two or more mRNA species or two or more cDNA species transcribed from said mRNA species, wherein said mRNA species or cDNA species are carried on a solid support, so as to determine the degree of correlation indicative of the presence of schizophrenia or a stage thereof, and so as to diagnose or identify schizophrenia or a stage thereof in said test human.   
     
     
         30 . The method as claimed in  claim 29 , wherein in step (a), the resulting isolated mRNA is subjected to reverse transcription to obtain isolated cDNA. 
     
     
         31 . The method as claimed in  claim 30 , where said isolated cDNA is amplified. 
     
     
         32 . The method as claimed in  claim 29 , wherein when isolated cDNA is obtained, any of said isolated cDNA is labeled. 
     
     
         33 . The method as claimed in  claim 29 , wherein in step (b), between 50 and 100 mRNA species or cDNA species are used. 
     
     
         34 . The method as claimed in  claim 29 , wherein, in step (b), between 10 and 500 mRNA species or cDNA species are used. 
     
     
         35 . The method as claimed in  claim 29 , wherein said solid support is a filter. 
     
     
         36 . The method as claimed in  claim 29 , wherein said method is used to detect a human's reaction to drug treatment during the treatment of schizophrenia.