Markers for prenatal diagnosis and monitoring

Abstract

Methods and kits are provided for diagnosing, monitoring, or predicting preeclaimpsia in a pregnant woman, trisomy 18 and trisomy 21 in a fetus, as well as for detecting pregnancy in a woman, by quantitatively measuring in the maternal blood the amount of one or more RNA species derived from a set of genetic loci and comparing the amount of the RNA species with a standard control.

1 . A method for diagnosing, monitoring, or predicting preeclampsia in a pregnant woman, the method comprising the steps of:
 (i) quantitatively determining the amount of one or more RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species are independently selected from RNA derived from genetic loci consisting of IGFBP3, ABP1, FN1, SLC21A2, KIAA0992, TIMP3, LPL, INHBA, LEP, ADAM12, PAPPA, PAPPA2, and SIGLEC6, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) comparing the amount of the RNA species from step (i) to a standard control representing the amount of the RNA species in the corresponding sample from an average non-preeclamptic pregnant woman, wherein an increase or a decrease in the amount of the RNA species from the standard control indicates preeclampsia or an increased risk of developing preeclampsia.   
     
     
         2 . The method of  claim 1 , wherein the RNA species is derived from ADAM12, PAPPA2, FN1, INHBA, LEP, or SIGLEC6, and wherein an increase in the amount of the RNA species from the standard control indicates preeclampsia or an increased risk of developing preeclampsia. 
     
     
         3 . The method of  claim 1 , wherein the RNA species is derived from PAPPA and wherein a decrease in the amount of the RNA species from the standard control indicates preeclampsia or an increased risk of developing preeclampsia. 
     
     
         4 . The method of  claim 1 , wherein step (i) comprises using a reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
         5 . The method of  claim 4 , wherein step (i) further comprises using mass spectrometry following RT-PCR. 
     
     
         6 . The method of  claim 1 , wherein step (i) comprises using a polynucleotide hybridization method. 
     
     
         7 . The method of  claim 1 , wherein step (i) comprises using a primer extension reaction. 
     
     
         8 . The method of  claim 1 , wherein the woman is during the first trimester of gestation. 
     
     
         9 . The method of  claim 1 , wherein the woman is during the second or third trimester of gestation. 
     
     
         10 . The method of  claim 1 , wherein the blood is plasma. 
     
     
         11 . The method of  claim 1 , wherein the blood is serum. 
     
     
         12 . The method of  claim 1 , wherein the increase in the amount of RNA from the standard control is more than 2-fold. 
     
     
         13 . The method of  claim 1 , wherein the decrease in the amount of RNA from the standard control is more than 50%. 
     
     
         14 . A kit for diagnosing, monitoring, or predicting preeclampsia in a pregnant woman, the kit comprising:
 (i) PCR primers for quantitatively determining the amount of one or more RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species is independently selected from RNA derived from genetic loci consisting of IGFBP3, ABP1, FN1, SLC21A2, KIAA0992, TIMP3, LPL, INHBA, LEP, ADAM12, PAPPA, PAPPA2, and SIGLEC6, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) a standard control representing the amount of the RNA species in the corresponding sample from an average non-preeclamptic pregnant woman.   
     
     
         15 . A method for detecting the presence of a fetus with trisomy 18 in a pregnant woman, the method comprising the steps of:
 (i) quantitatively determining the amount of an RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species is derived from RPL17, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) comparing the amount of the RNA species from step (i) to a standard control representing the amount of the RNA species in the corresponding sample from an average pregnant woman with a chromosomally normal fetus, wherein an increase in the amount of the RNA species from the standard control indicates an increased risk of having a fetus with trisomy 18.   
     
     
         16 . The method of  claim 15 , wherein step (i) comprises using reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
         17 . The method of  claim 16 , wherein step (i) further comprises using mass spectrometry. 
     
     
         18 . The method of  claim 15 , wherein step (i) comprises using a polynucleotide hybridization method. 
     
     
         19 . The method of  claim 15 , wherein step (i) comprises using a primer extension reaction. 
     
     
         20 . The method of  claim 15 , wherein the woman is during the first trimester of gestation. 
     
     
         21 . The method of  claim 15 , wherein the woman is during the second or third trimester of gestation. 
     
     
         22 . The method of  claim 15 , wherein the blood is plasma. 
     
     
         23 . The method of  claim 15 , wherein the blood is serum. 
     
     
         24 . The method of  claim 15 , wherein the increase in the amount of mRNA from the standard control is more than 2-fold. 
     
     
         25 . The method of  claim 15 , wherein the decrease in the amount of mRNA from the standard control is more than 50%. 
     
     
         26 . A kit for detecting the presence of a fetus with trisomy 18 in a pregnant woman, the kit comprising:
 (i) PCR primers for quantitatively determining the amount of an RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species is derived from RPL17, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) a standard control representing the amount of the RNA species in a corresponding sample from an average pregnant woman with a chromosomally normal fetus.   
     
     
         27 . A method for detecting the presence of a fetus with trisomy 21 in a pregnant woman, the method comprising the steps of:
 (i) quantitatively determining the amount of one or more RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species is independently selected from RNA species derived from genetic loci consisting of COL6A1, COL6A2, SOD1, APP, BTG3, ATP5J, ADAMTS1, BACE2,DSCR5, ITSN1, PLAC4, ATP5O, LOC90625, EFEMP1, and TFRC, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) comparing the amount of the RNA species from step (i) to a standard control representing the amount of the RNA species in the corresponding sample from an average pregnant woman with a chromosomally normal fetus, wherein an increase or a decrease in the amount of RNA species from the standard control indicates an increased risk of having a fetus with trisomy 21.   
     
     
         28 . The method of  claim 27 , wherein the RNA species is derived from ADAMTS1, APP, ATP5O, EFEMP1, or TFRC, and an increase in the amount of the RNA species from the standard control indicates an increased risk of having a fetus with trisomy 21. 
     
     
         29 . The method of  claim 27 , wherein step (i) comprises using a reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
         30 . The method of  claim 29 , wherein step (i) further comprises using mass spectrometry. 
     
     
         31 . The method of  claim 27 , wherein step (i) comprises using a polynucleotide hybridization method. 
     
     
         32 . The method of  claim 27 , wherein step (i) comprises using a primer extension reaction. 
     
     
         33 . The method of  claim 27 , wherein the woman is during the first trimester of gestation. 
     
     
         34 . The method of  claim 27 , wherein the woman is during the second or third trimester of gestation. 
     
     
         35 . The method of  claim 27 , wherein the blood is plasma. 
     
     
         36 . The method of  claim 27 , wherein the blood is serum. 
     
     
         37 . The method of  claim 27 , wherein the increase in the amount of the RNA species from the standard control is more than 2-fold. 
     
     
         38 . The method of  claim 27 , wherein the decrease in the amount of the RNA species from the standard control is more than 50%. 
     
     
         39 . A kit for detecting the presence of a fetus with trisomy 21 in a pregnant woman, the kit comprising:
 (i) PCR primers for quantitatively determining the amount of one or more RNA species in a biological sample obtained from the pregnant woman, wherein the RNA species is independently selected from RNA species derived from genetic loci consisting of COL6A1, COL6A2, SOD1, APP, BTG3, ATP5J, ADAMTS1, BACE2, DSCR5, ITSN1, PLAC4, ATP5O, LOC90625, EFEMP1, and TFRC, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) a standard control representing the amount of the RNA species in the corresponding sample from an average pregnant woman with a chromosomally normal fetus.   
     
     
         40 . A method for detecting pregnancy in a woman, the method comprising:
 (i) quantitatively determining the amount of one or more RNA species in a biological sample obtained from the woman, wherein the RNA species is independently selected from RNA species derived from genetic loci consisting of COL6A1, COL6A2, SOD1, ATP5O, ADAMTS1, DSCR5, and PLAC4, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) comparing the amount of the RNA species from step (i) to a standard control representing the amount of the RNA species in the corresponding sample from an average non-pregnant woman, wherein an increase or decrease in the amount of the RNA species from the standard control indicates pregnancy.   
     
     
         41 . The method of  claim 40 , wherein the RNA species is derived from COL6A1, COL6A2, ATP5O, or PLAC4, and wherein an increase in the amount of the RNA species from the standard control indicates pregnancy. 
     
     
         42 . The method of  claim 40 , wherein step (i) comprises using a reverse transcriptase polymerase chain reaction (RT-PCR). 
     
     
         43 . The method of  claim 42 , wherein step (i) further comprises using mass spectrometry. 
     
     
         44 . The method of  claim 40 , wherein step (i) comprises using a polynucleotide hybridization method. 
     
     
         45 . The method of  claim 40 , wherein step (i) comprises using a primer extension reaction. 
     
     
         46 . The method of  claim 40 , wherein the woman is during the first trimester of gestation. 
     
     
         47 . The method of  claim 40 , wherein the woman is during the second or third trimester of gestation. 
     
     
         48 . The method of  claim 40 , wherein the blood is plasma. 
     
     
         49 . The method of  claim 40 , wherein the blood is serum. 
     
     
         50 . The method of  claim 40 , wherein the increase in the amount of RNA from the standard control is more than 2-fold. 
     
     
         51 . The method of  claim 40 , wherein the decrease in the amount of RNA from the standard control is more than 50%. 
     
     
         52 . A kit for detecting pregnancy in a woman, the kit comprising:
 (i) PCR primers for quantitatively determining the amount of one or more RNA species in a biological sample obtained from the woman, wherein the RNA species is independently selected from RNA species derived from genetic loci consisting of COL6A1, COL6A2, SOD1, ATP5O, ADAMTS1, DSCR5, and PLAC4, and wherein the biological sample is blood, washing from the reproductive tract, amniotic fluid, urine, saliva, or chorionic villus; and   (ii) a standard control representing the amount of the RNA species in the corresponding sample from an average non-pregnant woman.