The Importance of Molecular Biological Analysis for the Laboratory Diagnostic of Homozygous Haemoglobin Malay.

Haemoglobin (Hb) Malay is variant haemoglobin with a ß++ thalassemia phenotype. The prevalence of Hb Malay in the Malaysian population was 5.5%. We describe a 58-year-old male who presented with symptomatic anaemia to the Hospital Universiti Sains Malaysia. Further history revealed that the patient had anaemia since the age of 28, and on regular follow-up at other hospital. Physical examination revealed pallor, jaundice and hepatosplenomegaly. The full blood count and peripheral blood smear showed hypochromic microcytic anaemia with anisopoikilocytosis, and many target cells. High-performance liquid chromatography results showed a ß thalassemia trait. However, the diagnosis does not alight with the patient's condition. Bone marrow aspirate was completed and showed reactive changes and erythroid hyperplasia. A molecular test was then performed for ß globin gene mutation detection using Multiplex Amplification Refractory Mutation System (M-ARMS) PCR method. This revealed the result as homozygous codon 19 mutation or Hb Malay. Therefore, in this case report we would like to highlight the laboratory approaches, the challenges faced by the usual haematological investigations and the importance role of molecular testing in the diagnosis of severe anaemia.

Clinical implications of conventional cytogenetics, fluorescence in situ hybridization (FISH) and molecular testing in chronic myeloid leukaemia patients in the tyrosine kinase inhibitor era - A review.

Chronic myeloid leukaemia (CML) provides an illustrative disease model for both molecular pathogenesis of cancer and rational drug therapy. Imatinib mesylate (IM), a BCR-ABL1 targeted tyrosine kinase inhibitor (TKI) drug, is the first line gold standard drug for CML treatment. Conventional cytogenetic analysis (CCA) can identify the standard and variant Philadelphia (Ph) chromosome, and any additional complex chromosome abnormalities at diagnosis as well as during treatment course. Fluorescence in situ hybridization (FISH) is especially important for cells of CML patients with inadequate or inferior quality metaphases or those with variant Ph translocations. CCA in conjunction with FISH can serve as powerful tools in all phases of CML including the diagnosis, prognosis, risk stratification and monitoring of cytogenetic responses to treatment. Molecular techniques such as reverse transcriptase-polymerase chain reaction (RT-PCR) is used for the detection of BCR-ABL1 transcripts at diagnosis whereas quantitative reverse transcriptase-polymerase chain reaction (qRTPCR) is used at the time of diagnosis as well as during TKI therapy for the quantitation of BCR-ABL1 transcripts to evaluate the molecular response and minimal residual disease (MRD). Despite the excellent treatment results obtained after the introduction of TKI drugs, especially Imatinib mesylate (IM), resistance to TKIs develops in approximately 35% - 40% of CML patients on TKI therapy. Since point mutations in BCR-ABL1 are a common cause of IM resistance, mutation analysis is important in IM resistant patients. Mutations are reliably detected by nested PCR amplification of the translocated ABL1 kinase domain followed by direct sequencing of the entire amplified kinase domain. The objective of this review is to highlight the importance of regular and timely CCA, FISH analysis and molecular testing in the diagnosis, prognosis, assessment of therapeutic efficacy, evaluation of MRD and in the detection of BCR-ABL1 kinase mutations which cause therapeutic resistance in adult CML patients.

Factors influencing post-cryopreserved CD34+ cells viability in the harvested products of autologous haematopoietic stem cells.

The cryopreservation process of stem cells potentially cause the loss of CD34+ cells. The aim of this study is to evaluate association of patient, graft and technical characteristics with post cryopreserved CD34+ cells viability among lymphoproliferative disease namely multiple myeloma (MM) and lymphoma patients at Hospital Universiti Sains Malaysia (USM). This retrospective study was conducted in the Transplant Unit. A search of the hospital data (2008-2018) to identify 132 patients for both MM and lymphoma who underwent autologous peripheral blood haematopoietic stem cells (APBSC) mobilisation, and were successfully harvested and cryopreserved. Selected patients' profile as well as selected parameters of stem cell mobilization and cryopreservation were obtained from laboratory information system (LIS), record unit and the Transplant Unit. Multiple logistic regression (MLR) was used to find significant associated factors and P<0.05 was considered significant. The mean age of the patients was 39 years old with almost equal gender distribution and majority were lymphoma patients, 96 (72.7%) while 36 (27.3%) were multiple myeloma (MM) patients. The significant influencing factors of post-cryopreserved CD34+ cells viability were pre-cryopreserved CD34+ cell viability, total nucleated cells (TNC), and anti-platelet and antibiotics usage. Patients who are not on anti-platelet and have higher pre-cryopreserved CD34+ cells viability have higher chance for good post-cryopreserved CD34+ cells viability. While, those patients with higher TNC and on antibiotics have lower chance for good post cryopreserved CD34+ cells viability. This study showed patients who are not on anti-platelet and antibiotics will have higher probability of achieving good post cryopreserved CD34+ cells viability. The APBSC products with higher pre-cryopreserved CD34+ cells viability and lower TNC will achieve better post-cryopreserved CD34+ cells viability. The addition of extra plasma to the APBSC products is recommended to reduce the TNC.

Hypercalcaemic crisis as the presenting manifestation of abdominal tuberculosis: a case report.

This report deals with a young man having prolonged fever presenting with hypercalcaemic crisis. Subsequent investigations confirmed tuberculosis (TB) peritonitis in the absence of pulmonary involvement as the cause of his symptoms. His hypercalcaemia and fever resolved with anti-TB therapy. Abdominal TB needs to be included in the differential diagnosis of otherwise unexplained hypercalcaemia especially in our region where TB is an endemic problem and is treatable.