Examination of stability of bone marrow blood RNA in the PAXgene tube.
The PAXgene RNA blood collection tube is used for RNA of peripheral blood (PB) and the stability of PB RNA in this tube has already been reported. However, the stability of bone marrow blood (BM) RNA in the PAXgene tube is unknown. Thus, we examined the stability of BM RNA in the PAXgene tubes. BM from leukemia patients was collected into PAXgene and EDTA tubes and stored at 4 degrees C for 5 days. RNA isolated from both tubes was analyzed by a quantitative reverse-transcription polymerase chain reaction (RT-PCR) analysis. Porphobilinogen deaminase (PBGD) mRNA of BM (as high-expression mRNA) and Wilms tumor suppressor (WT1) mRNA of BM (as low-expression mRNA) and very low copies of major BCR-ABL mRNA (as minimal residual disease of leukemia) of leukemia of BM were quantified by a LightCycler system. RNA yield from the PAXgene tubes and the intensity of 28S rRNA bands on RNA electrophoresis showed a degradation trend. However, the intensity of 18S rRNA bands from the PAXgene tubes remained. The expression of PBGD and WT1 of BM in the PAXgene tubes did not decrease for 5 days. The very low copies of major BCR-ABL mRNA in the PAXgene tubes were detectable on day 5 but those in the EDTA tubes were not detectable. Therefore, the PAXgene tube can be used for BM samples in a quantitative RT-PCR of the fusion gene transcripts of leukemia.
Yamamoto
Toshiharu
T
The Fourth Division of Clinical Investigation, Kanagawa Cancer Center, Nakao 1-1-2 Asahi Ward, Yokohama City, Japan 241-0815. kccmolecularbio@hotmail.com
Sekiyama
Akiko
A
Sekiguchi
Hironobu
H
Yoshida
Tsutomu
T
Miyagi
Yohei
Y
D001800:Blood Specimen Collection / Q000295:instrumentation* / Q000379:methods*; D001854:Bone Marrow Cells / Q000378:metabolism*; D001856:Bone Marrow Examination / Q000379:methods; D006801:Humans; D007938:Leukemia / Q000235:genetics; D020871:RNA Stability; D012333:RNA, Messenger / Q000032:analysis*; D012334:RNA, Neoplasm / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D012334:RNA, Neoplasm