FISH versus real-time quantitative PCR for monitoring of minimal residual disease in chronic myeloid leukaemia patients on tyrosine kinase inhibitor therapy.

INTRODUCTION: BCR-ABL fusion gene, the oncogenic driver of CML, results from a translocation between short arms of chromosome 9 and 22. Monitoring of CML patients during treatment is essential, not only for tailoring the treatment but also to detect early relapse to enable timely intervention. Commonly used methods for detection of residual disease are conventional karyotyping, FISH and molecular methods. In this study, we compared FISH with QRT-PCR for detection of residual disease in CML. MATERIALS AND METHODS: CML patients on tyrosine kinase inhibitor (TKI) therapy and on regular follow up at University Kebangsaan Malaysia Medical Center (UKMMC) were selected. A comparative study was conducted between FISH and QRT-PCR for BCR-ABL transcripts at diagnosis and during follow-up. RESULTS: There was good correlation between FISH and QRT-PCR for BCR-ABL. At 6th month of follow-up post diagnosis, FISH had a sensitivity of 83.3% and specificity of 65.2% (k >0.339, p<0.033). At 12th month, the sensitivity of FISH was 83% and the specificity was 59.1% (k >0.286, p <0.065). Similarly, at the 24th month, FISH had a sensitivity of 100% and specificity of 68.8% (k >0.642, p<0.000). DISCUSSION: Early achievement of major molecular response (MMR) and complete cytogenetic remission (CCyR) were reliable predictors of long-term maintenance of molecular remission.

Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis.

INTRODUCTION: Haemoglobin Bart's (Hb Bart's) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart's using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). METHODS: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart's by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the ß-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. RESULTS: This study was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart's peak using the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods. The range of Hb Bart's using NF programme and CB programme were (0.5-4.1%) and (0.5-7.1%), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7 and three αα/ααCS. Fifty Hb Bart's negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of 100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. CONCLUSION: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but acceptable sensitivities in the detection of Hb Bart.

Tunable single Stokes extraction from 20 GHz Brillouin fiber laser using ultranarrow bandwidth optical filter.

The individual extraction of a Brillouin Stokes line from a 20 GHz comb generated from the compact configuration of a multiwavelength Brillouin fiber ring laser configuration has been achieved using an ultranarrow bandwidth (UNB) optical filter. The narrowest bandwidth transmission of a UNB optical filter that is 50 pm is used in order to get particular Stokes. The Stokes filtered is in the wavelength range of 1549.768-1551.016 nm. High SNR within the range of 54.97-11.73 dB with almost nil peak power loss being obtained was monitored by a 0.16 pm optical spectrum analyzer, giving convincing results. Relatively, the proposed configuration could provide wide tunability and narrow selection of the Brillouin Stokes.

Closely spaced dual-wavelength fiber laser using an ultranarrow bandwidth optical filter for low radio frequency generation.

A dual-wavelength (DW) fiber laser with a closely spaced single longitudinal mode (SLM) output is proposed and demonstrated. The proposed fiber laser utilizes a conventional fiber Bragg grating with a center wavelength of about 1546.8 nm in conjunction with an ultranarrow bandwidth tunable optical filter to generate the desired DW SLM output. Observations with a very high resolution optical spectrum analyzer, which was capable of achieving resolutions up to 0.16 pm, revealed detailed spectral characteristics not characteristically seen before. A channel spacing of up to 58 nm was realized, and spacing as small as 2 pm was achieved. The minimum channel spacing and its resulting beat frequency are the narrowest observed yet to the best of our knowledge for a DW fiber laser at room temperature.