single nucleotide polymorphism (snp)-based loss of heterozygosity (loh) testing by real time pcr in patients suspect of myeloproliferative disease单核苷酸多态性(snp)的杂合性丢失(loh)的实时pcr检测患者骨髓增殖性疾病的怀疑.pdf

Single Nucleotide Polymorphism (SNP)-Based Loss of Heterozygosity (LOH) Testing by Real Time PCR in Patients Suspect of Myeloproliferative Disease 1 1 2 2 Cornelis J. J. Huijsmans *, Jeroen Poodt , Jan Damen , Johannes C. van der Linden , 3 4 5 1 Paul H. M. Savelkoul , Johannes F. M. Pruijt , Mirrian Hilbink , Mirjam H. A. Hermans 1 Laboratory of Molecular Diagnostics, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands, 2 Laboratory of Pathology, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands, 3 Medical Microbiology and Infection Control, Vrije Universiteit (VU) University Medical Center, Amsterdam, The Netherlands, 4 Department of Internal Medicine, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands, 5 Jeroen Bosch Academy, Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands Abstract During tumor development, loss of heterozygosity (LOH) often occurs. When LOH is preceded by an oncogene activating mutation, the mutant allele may be further potentiated if the wild-type allele is lost or inactivated. In myeloproliferative neoplasms (MPN) somatic acquisition of JAK2V617F may be followed by LOH resulting in loss of the wild type allele. The occurrence of LOH in MPN and other proliferative diseases may lead to a further potentiating the mutant allele and thereby increasing morbidity. A real time PCR based SNP profiling assay was developed and validated for LOH detection of the JAK2 region (JAK2 LOH). Blood of a cohort of 12 JAK2V617F-positive patients (n = 6 25–50% and n = 6.50% JAK2V617F) and a cohort of 81 patients suspected of MPN was stored with EDTA and subsequently used for validation. To generate ger