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Simultaneous detection of JAK2, CALR, and MPL mutations and quantitation of JAK2 V617F allele burden in myeloproliferative neoplasms using the quenching probe-Tm method in i-densy IS-5320.
Arai, Kunihito; Sakaguchi, Masahiro; Yui, Shunsuke; Kitano, Tomoaki; Miyata, Miho; Yogosawa, Mayumi; Nakayama, Kazutaka; Tajika, Kenji; Usuki, Kensuke; Kuroda, Junya; Uoshima, Nobuhiko; Kobayashi, Yutaka; Uchiyama, Hitoji; Kubota, Yasushi; Kimura, Shinya; Mori, Shinichiro; Hirai, Mitsuharu; Wakita, Satoshi; Yamaguchi, Hiroki.
Afiliação
  • Arai K; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Sakaguchi M; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Yui S; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Kitano T; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Miyata M; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Yogosawa M; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Nakayama K; Department of Hematology, Yokohama Minami Kyousai Hospital, Kanagawa, Japan.
  • Tajika K; Department of Hematology, Yokohama Minami Kyousai Hospital, Kanagawa, Japan.
  • Usuki K; Department of Hematology, NTT Medical Center Tokyo, Tokyo, Japan.
  • Kuroda J; Division of Hematology and Oncology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
  • Uoshima N; Department of Hematology, Japanese Red Cross, Kyoto Daini Hospital, Kyoto, Japan.
  • Kobayashi Y; Department of Hematology, Japanese Red Cross, Kyoto Daini Hospital, Kyoto, Japan.
  • Uchiyama H; Department of Hematology, Japanese Red Cross Kyoto Daiichi Hospital, Kyoto, Japan.
  • Kubota Y; Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, Saga, Japan.
  • Kimura S; Division of Hematology, Respiratory Medicine and Oncology, Department of Internal Medicine, Faculty of Medicine, Saga University, Saga, Japan.
  • Mori S; Hemato-Oncology Department, St Luke's International Hospital, Tokyo, Japan.
  • Hirai M; ARKRAY, Inc., Kyoto, Japan.
  • Wakita S; Department of Hematology, Nippon Medical School, Tokyo, Japan.
  • Yamaguchi H; Department of Hematology, Nippon Medical School, Tokyo, Japan.
Int J Lab Hematol ; 44(6): 1102-1110, 2022 Dec.
Article em En | MEDLINE | ID: mdl-36039795
ABSTRACT

INTRODUCTION:

Accurate detection of myeloproliferative neoplasms (MPN)-associated gene mutations is necessary to correctly diagnose MPN. However, conventional gene testing has various limitations, including the requirement of skilled technicians, cumbersome experimental procedures, and turnaround time of several days. The gene analyzer i-densy IS-5320 allows gene testing using the quenching probe-Tm method. Specifically, pretreatment of samples including DNA extraction, amplification and detection of genes, and analysis of results are performed in a fully automatic manner after samples and test reagents are added into this system, which is compact and can be easily installed in a laboratory. The aim of this study is to investigate the sensitivity and specificity associated with the simultaneous detection of MPN-associated gene mutations.

METHODS:

We conducted an analysis of MPN-associated genes using i-densy IS-5320. We analyzed 384 samples (171 JAK2 V617F mutations, 10 JAK2 exon12 mutations, 104 CALR mutations, and 26 MPL mutations) that had been examined using conventional approaches such as allele-specific polymerase chain reaction (PCR), droplet digital PCR, and the direct sequencing method.

RESULTS:

The detection accuracy of JAK2 V617F, JAK2 exon 12, CALR, and MPL was 100.0% (383/383), 99.7% (383/384), 100.0% (370/370), and 99.7% (377/378), respectively. There was a strong positive correlation between the JAK2 V617F allele burden measured using conventional methods and i-densy IS-5320 (r = .989).

CONCLUSION:

Overall, i-densy IS-5320 exhibited good accuracy in terms of analyzing MPN-associated genes; thus, it can serve as a replacement for conventional methods of MPN-associated gene testing.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transtornos Mieloproliferativos / Neoplasias Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transtornos Mieloproliferativos / Neoplasias Tipo de estudo: Diagnostic_studies Limite: Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article