Individualized PCR strategies hamper comparability of molecular results between different laboratories in several fields of medicine. To harmonize BCR-ABL mRNA quantification an international multicenter trial involving 37 laboratories in 14 countries was initiated using 10 samples, each containing various dilutions (10, 2, 1 and 0.1%) of b3a2 or b2a2 BCR-ABL positive in normal leukocytes and negative controls. A novel control plasmid (pME-2) was designed for external calibration containing BCR-ABL and glucuronidase-b (GUS) sequences. Median BCR-ABL/ABL ratios were 9.1, 1.8, 0.85 and 0.11% in b3a2 samples and 9.5, 1.6, 0.84 and 0.11% in b2a2 samples. Median BCR-ABL/GUS ratios were 3.4, 0.77, 0.37 and 0.042% in b3a2 samples and 2.8, 0.48, 0.29 and 0.031% in b2a2 samples. The coefficients of variation were 0.62 for ratios BCR-ABL/ABL and 1.03 for ratios BCR-ABL/GUS. Five of 37 evaluable participating laboratories (13%) detected low BCR-ABL copy numbers in negative control samples; one laboratory failed to detect BCR-ABL in a low-level sample. We conclude that the use of a common control plasmid does indeed improve comparability of BCR-ABL mRNA quantification results. However, further standardizing efforts like introducing a calibrator and regular control rounds are needed.
Summary Background and Methods: As a source of hematopoietic stem cells, cord blood (CB) is an alternative to bone marrow or peripheral blood stem cells (PBSC). The Mannheim CordBlood Bank has currently stored about 1,750 allogeneic CB units. Here we report our experiences and discuss future perspectives of CB banking. We analyzed CB units for nucleated cell (NC), mononucleated cell (MNC) and CD34+ cell count, volume, colony-forming units (CFU-GM) as well as ethnic background of the donor. Transplanted CB units were analyzed for patient and transplant characteristics and compared to stored CB units. Results: Only 25% of all collected CB units met storage criteria. Main reasons for exclusion were: i) insufficient volume (57.7%), ii) delayed arrival at the processing site (19.2%) and iii) little cell count (7.2%). Up to now 36 CB units have been released for transplantation mainly to children (62%). Transplant indications were hematological diseases, immune deficiencies and metabolic diseases. Transplanted CB units showed significantly higher cell counts compared to stored units (NC: 12.5 vs. 7.2 × 10 8 , MNC: 4.7 vs. 2.9 × 10 8 , CD34+ cells: 3.3 vs. 1.8 × 10 6 , mean; p < 0.001 each) and were found more often in ethnic minority groups (36 vs. 20%; p = 0.04). Conclusions: Even though cell count and volume are key parameters for the eligibility of CB units, our data indicate that the ethnic background of the donor also plays a major role. Collection and processing of CB should be optimized in order to gain maximum volume and cell counts.Schlüsselwörter Nabelschnurblut · Nabelschnurblut-Aufarbeitung · Stammzelltransplantation · Hämatopoetische Stammzellen Zusammenfassung Hintergrund und Methoden: Nabelschnurblut (CB) ist eine Alternative zur Gewinnung von hämatopoetischen Stammzellen aus Knochenmark oder peripheren Stammzellen. Die Mannheimer Nabelschnurblutbank hat bisher 1750 allogene Nabelschnurblutpräparate eingelagert. Wir berichten über unsere Erfahrungen und diskutieren Zukunftsperspektiven der Nabelschnurbluteinlagerung. Folgende CB-Parameter wurden untersucht: Volumen, nukleäre (NC), mononukleäre (MNC) und CD34+ Zellen, Kolonie bildende Einheiten (CFU-GM) sowie ethnische Herkunft des Spenders. Bei den transplantierten Präparaten wurden die Patienten-und Transplantatdaten ausgewertet und die Präparateeigenschaften mit eingelagerten CB verglichen. Ergebnisse: Nur 25% aller gesammelten Nabelschnurblutpräparate erfüllten die Einlagerungskriterien. Die Hauptursachen hierfür waren 1) zu niedriges Vollblutvolumen (57,7%), 2) zu spätes Eintreffen der Prä-parate im Verarbeitungszentrum (19,2%) sowie 3) zu niedrige Zellzahl (7,2%). Bisher wurden 36 Mannheimer Nabelschnurblutpräparate, vorwiegend an Kinder (62%) abgegeben. Die Transplantationsindikationen waren primär hämatologische Erkrankungen, Erkrankungen des Immunsystems und metabolische Erkrankungen. Die Zellzahlen der transplantierten Präparate waren signifikant höher, als die der eingelagerten (NC: 12,5 vs. 7,2 × 10 8 , MNC: 4,7 vs. 2,9 × 10 8 , CD34+ Zellen:...
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