Johnsen_EU Haematology 03/03/2010 13:12 Page 40
Haematological Malignancies
Table 1: Clinical Data for Adult Patients from Conclusive Multiplex Reverse Transcriptase
Polymerase Chain Reaction
Adult Patients with Adult Patients Diagnosed Paediatric Patients with
Multiplex RT-PCR 1/1/1992 – 31/12/2003 Multiplex RT-PCR Performed
Performed (n=191) (n=329) (n=11)
Age (years) Median 66 67 4
Range 24–89 22–91 0–14
IQ range 54–74 56–75 1–12
% <55 28 24 100
Male/female ratio 1.17 1.23 1.75
FAB classification M0 8 9 1
M1 35 61
M2 38 71 2
M3 7 9
M4 37 62 5
M5 14 14 1
M6 1 6 2
M7 3 9
Not classified 48 89
% blast cells in BM Median 59 50 56
Range 1–100 1–100 11–90
IQ range 32–81 28–76 15–80
WBCs in median peripheral blood Median 14.6 10.0 18.6
Range 0.7–443.2 0.2–443.2 1.8–450.0
IQ range 3.4–52.8 2.3–42.3 5.5–52.0
CR rate* 66.0% (62/94) 59.9% (91/152) 90.9% (10/11)
Median survival in years Median 0.4 0.4 ND
Range 0.0–9.1 0.0–10.6
IQ range* 0.1–1.1 0.1–1.1
Conclusive cytogenetics available 132 (69%) 188 (57%) 11 (100%)
Clinical data for adult patients with conclusive multiplex reverse transcriptase polymerase chain reaction (RT-PCR) performed, for total number of adult patients diagnosed during the
inclusion period and for paediatric patients with conclusive multiplex RT-PCR performed.
*Number of patients achieving complete response/number of patients with curative treatment intention.
BM = bone marrow; CR = complete response; FAB = French–American–British; IQ = interquartile; ND = not declared; WBCs = white blood cells.
The RT-PCR technology has the advantage of being independent of the study group of 191 samples analysed (see Table 1). In addition, 11
dividing cells. Furthermore, RT-PCR allows identification of the vast paediatric patients with AML were included prospectively at the time of
majority of patients with t(8;21), inv(16) or t(15;17), which defines the diagnosis. This patient material has been published previously.
1
group of patients with relatively favourable treatment outcome.
14–16
In
addition, in fusion-gene-positive cases, a marker for minimal residual RNA Extraction, Reverse Transcription and
disease monitoring is identified. Multiplex Polymerase Chain Reaction
Total RNA was extracted using the QIAamp
®
RNA Blood Mini Kit
In terms of the establishment of an overall consensus concerning gene (QIAGEN, Merck Euro lab A/S, Albertslund, Denmark) according to the
diagnostics in haematology, we performed an HTA using retrospective manufacturer’s recommendations. Analyses for balanced translocations
and prospective data collected from an accessible regional clinical were performed by multiplex RT-PCR analysis using the HemaVision
database and biobank. screen test according to the manufacturer’s recommendations.
17
Materials and Method The commercially available multiplex RT-PCR assay detects 28
Trial Design and Approval different chromosomal abnormalities by testing for the presence of
This HTA focused on aspects of the technology, the patient, the more than 80 fusion transcript variants. Reverse transcription is
organisation and economic factors related to the introduction of a performed with a mixture of translocation-specific primers. After
commercial multiplex RT-PCR screening assay covering 28 leukaemia complementary DNA (cDNA) synthesis, the PCR amplification is
fusion transcripts (HemaVision
®
; DNA Technology, Aarhus, Denmark) performed in two steps: a master PCR amplification followed by
identifying favourable risk translocations in AML. The programme was nested PCR, which screens for the presence of fusion transcripts, and
accepted by the local ethics committee in Copenhagen. a split-out PCR amplification followed by nested PCR, which identifies
the specific fusion transcript(s).
Patients and Methods
A total of 329 adult patients (>15 years of age) were diagnosed with AML Cytogenetics
or chronic myeloid leukaemia (CML) blast crisis at a single institution Cytogenetic analysis was reported at diagnosis in 143 of the cases
during a 12-year period between 1 January 1992 and 31 December in which conclusive multiplex RT-PCR was performed. Clonal
2003. Clinical and biological information was collected from patient files abnormalities were defined in accordance with the International
for all adult patients with information available (n=321, 98%) to define System for Human Cytogenetic Nomenclature.
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EUROPEAN HAEMATOLOGY
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