Prenatal Screening of Chromosomal Abnormalities in Fetal DNA Circulating in Maternal Blood

KamTek is supporting the development of HarmonyTM Prenatal Test for chromosomal abnormalities in fetal DNA circulating in mother’s blood.  This is accomplished by applying NextGen sequencing techniques detecting DNA sequences specific for each of the abnormality. The diagnostic kits are being developed by Roche with clinical tests performed at Ariosa diagnostics, the inventor of the technique.  Control DNAs used in these test kits come from specific EBV transformed cell lines developed at KamTek, Inc.

KTI worked with Ariosa Diagnostics to EBV-transform B-cells from small quantities of cord blood from early stage pregnancy to establish cell lines for specific chromosomal abnormalities. First such successfully established cell line is Trisomy 18 from cord blood along with normal cell line from the mother blood to yield large quantities of DNA of each type. This service provided to Ariosa Daignostics has become the basis of a variety of prenatal “Harmony test kits” developed in alliance with Roche sequencing Inc. The HarmonyTM Prenatal Test is a blood screening test for pregnant women that can be used as early as 10 weeks of pregnancy to evaluate cell-free fetal DNA circulating in maternal blood and diagnose the presence of this T18 trisomy (down syndrome) by NextGen Sequencing analyses.

Following steps are taken for this project:

• Receive Paired Mother and Cord blood samples, 
• Perform EBV transformation on both samples
• Expand cultures
• Cryo-preserve early passages during expansion
• Also Perform cytogenetic testing during expansion on a small sample of the master culture
• Continue expansion and cryo-preservations if cord cultures show the expected chromosomal abnormality
• Perform sterility and Mycoplasma test on the expanded culture
• Continue expansion and collect cells in a pellets for DNA isolations
• Perform DNA isolations
• Submit cell pellets and isolated DNA to Roche for further evaluation
• Continue expansion, cryo-preservations and Cell pellets preparations per requirement

Roche Sequencing Solutions Inc. has expanded such blood test to diagnose a variety of other chromosomal abnormalities from circulating fetal DNA in mother’s blood. KamTek continues to assist Roche sequencing Inc. for  providing cellular resource development of  HarmonyTM kit for this T18 trisomy as well as for development of  additional cell lines via EBV transformation for other abnormalities. Currently KamTek is engaged in development of cell lines specific for T21 trisomy. Roche is expanding the use of this prenatal test for global diagnostic market using KamTek as its valuable cell line supplier.