Multiple myeloma sequencing in BIOMED-2 PCR using cDNA as template

Shimada Shotaro, Kuroiwa Kai, Narita Hinako, Okamura Reiko, Uesugi Yuka, Sasaki Yohei, Watanuki Megumi, Arai Nana, Kawaguchi Yukiko, Fujiwara Shun, Yanagisawa Koji, Hattori Norimichi

doi:10.14930/jshowaunivsoc.84.10

Detection technology for measurable residual disease in multiple myeloma （MM） has improved over the years. High detection sensitivity for polymerase chain reaction （PCR）-based identification of immunoglobulin （Ig） region has been obtained using allele-specific oligonucleotide primer; however, certain limitations regarding the complexity of Ig region sequencing or specific PCR primer designing still remain. We herein performed BIOMED-2 PCR, which was developed in 2003, using the RNA extracted from bone marrow samples of patients with MM as a template and confirmed the success rate of Ig region decoding via sequencing after TA-cloning. BIOMED-2 PCR, performed using cDNA as a template, demonstrated a success rate of 58.1％ with IHGV-J-specific primers. Further, univariate and multivariate analyses indicated significantly higher PCR success rates in IgG and IgA types than Bence Jones protein （BJP） type. The low success rate in the BJP type may be due to reduced RNA expression and abnormal IgH reconstitution. BIOMED-2 PCR was effectively performed using cDNA that was prepared from the RNA sample extracted from long-term stored specimens, regardless of plasma cell percentage during specimen collection or nucleic acid concentration during extraction.

Multiple myeloma sequencing in BIOMED-2 PCR using cDNA as template

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