Analysis of class I and II aberrations in Iraqi childhood acute myeloid leukemia using filter paper cards

Abstract

The lack of molecular diagnosis in the field of cancer in Iraq has motivated us to perform a genetic analysis of pediatric acute myelogenous leukemia (AML), including class I and II aberrations. Peripheral blood or bone marrow cells were collected from 134 AML children aged ≤15 years. Flinders Technology Associates (FTA) filter paper cards were used to transfer dried blood samples from five Iraqi hospitals to Japan. DNA sequencing was performed to identify class I mutations. Nested RT-PCR was used to detect class II aberrations, except that MLL rearrangement was detected according to long distance inverse-PCR. NPM1 and FMS-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutations were analyzed by GeneScan using DNA template. Among 134 Iraqi pediatric AML samples, the most prevalent FAB subtype was M2 (33.6 %) followed by M3 (17.9 %). Class I mutations: 20 (14.9 %), 8 (6.0 %), and 8 (6.0 %) patients had FLT3-ITD, FLT3-TKD, and KIT mutations, respectively. Class II mutations: 24 (17.9 %), 19 (14.2 %), and 9 (6.7 %) children had PML-RARA, RUNX1-RUNX1T1, and CBFB-MYH11 transcripts, respectively. MLL rearrangements were detected in 25 (18.7 %) patients. NPM1 mutation was detected in seven (5.2 %) cases. Collectively, approximately 30 % of AML children were proved to carry favorable prognostic genetic abnormalities, whereas approximately 10 % had high FLT3-ITD allelic burden and needed a special treatment plan including allogeneic hematopoietic stem cell transplantation. Acute promyelocytic leukemia (APL) was frequent among Iraqi pediatric AML. It is likely that molecular diagnosis using FTA cards in underdeveloped countries could guide doctors towards an appropriate treatment strategy.

Analysis of KRAS and NRAS Gene Mutations in Arab Asian Children With Acute Leukemia: High Frequency of RAS Mutations in Acute Lymphoblastic Leukemia

“…Diagnostically difficult cases were reviewed by an Italian team guided by Dr. Anna Maria Testi (Department of Cellular Biotechnologies and Hematology, University “La Sapienza”, Rome, Italy) via telemedicine sessions . The diagnosis of ALL and AML was confirmed on initial unstained bone marrow (BM) aspiration smears that were transferred from Iraq and stained with May‐Grünwald‐Giemsa and myeloperoxidase at the Department of Pediatrics, Shinshu University School of Medicine . In Jordan, the diagnosis of ALL and AML was confirmed by flow cytometric analysis: CD10, CD19, CD20, CD22, and cytoplasmic CD79a were used for B‐precursor ALL; CD1a, CD2, CD3, CD4, CD5, CD7, CD8, cytoplasmic CD3, and TdT were used for T‐lineage ALL.…”

“…A total of 318 patients with ALL consisted of 264 previously reported ALL cases from five pediatric oncology centers in Iraq: the Children's Welfare Teaching Hospital in Baghdad (a major referral center for childhood cancers in the country), Central Teaching Hospital for Children in Baghdad, Basra Children's Specialty Hospital in Basra, Ibn Al‐Atheer Hospital for Children in Mosul, and Nana‐Kali Hospital for Hemato‐Oncology in Erbil, and 54 patients from KHCC in Jordan. A total of 167 patients with AML consisted of 134 previously reported Iraqi AML cases and 33 from KHCC were evaluated . All Iraqi samples and Jordanian ALL samples were collected using Flinders Technology Associates (FTA) cards between October 2009 and December 2012.…”

“…As reported previously, a few drops of diagnostic BM aspirate or PB were applied to FTA classic card matrices (Cat. No.WB120205, GE Healthcare UK Ltd., Buckinghamshire, UK) at the enrolled hospitals in Iraq and Jordan.…”

DOCK8 mutation diagnosed using whole-exome sequencing of the dried blood spot-derived DNA: a case report of an Iraqi girl diagnosed in Japan

“…The use of WES of the DBS-derived-DNA was the best approach to reach the diagnosis in our case. We previously utilized FTA cards to transfer samples of Iraqi children with acute leukemia and other hematological diseases [13][14][15][16].…”

“…Whole-exome sequencing (WES) is an advanced approach, covers > 95% of the exons which harbor most of the genetic variants associated with phenotypes of human diseases [12]. Likewise, Flinders Technology Associates (FTA) cards are convenient for dried blood spots (DBS) archiving, transportation, DNA/RNA extraction and further genetic analysis [13][14][15][16].…”