Year: 2008
Type: article
Study design: Observational Study
Abstract: Higher detection rate of JAK2 mutation using plasmaWe recently reported that using plasma rather than peripheral blood cells improves detection of the JAK2 V617F mutation, 1 which is the most important characteristic molecular abnormality in patients with nonchronic myeloid leukemia (non-CML)myeloproliferative disease (MPD).[2][3][4] Here we report the findings after expanding on our study and testing 634 consecutive paired plasma and cell samples for the V617F mutation and mutations in JAK2 exon 12.All 634 samples were collected from patients who were clinically suspected of having a non-CML MPD.We report that the V617F mutation was detected in 151 (24%) patients when plasma was used and only in 140 (22%) paired cell samples.No patients were positive by cell analysis and negative by plasma analysis while 7% of patients positive by plasma were negative by cells.When direct sequencing is used, a dominant (Ͼ 80%) mutant (T) peak is usually indicative of homozygous (2 mutant genes) or hemizygous mutation (one mutant gene and one deleted gene). 1 Of the 151 V617F-positive samples, 58 (38%) showed homozygous/ hemizygous mutation by plasma analysis; but only 36 (24%) were homozygous/hemizygous by cell analysis.All testing for JAK2 mutations was performed using direct bidirectional sequencing of JAK2 mRNA.Two PCR primers were designed to amplify a 273-bp segment of JAK2 encompassing the codon for amino acid 617.A second set of primers was designed to amplify a 491-bp segment encompassing exons 12, 13, and 14.The primer sequences used were as follows: 5Ј-GAC TAC GGT CAA CTG CAT GAA A-3Ј and 5Ј-CCA TGC CAA CTG TTT AGC AA-3Ј for the 273-bp segment and 5Ј-tgt aaa acg acg gcc agt CTA AAT GCT GTC CCC CAA AG-3Ј; and 5Ј-cag gaa aca gct atg acc CCA TGC CAA CTG TTT AGC AA-3Ј (uppercase indicates the actual sequences, and lower case represents the M13 linker) for the 491-bp.The Surveyor
Source: Blood
Language: English
Subfield: Genetics
Field: Medicine
Domain: Health Sciences
Open Access status: bronze
OpenAlex ID: W2094799752